Patent Publication Number: US-2011060574-A1

Title: Computer-based modeling and designing of phosphofructokinase 1 (pfk) modulators

Description:
The subject matters of the invention are: a crystallographic model of the binding site and a modulator regulating the catalytic activity of phosphofructokinase (PFK), a method of designing, selecting and producing a PFK modulator, a computer-based method for the analysis of the interaction between the modulator and PFK, a computer-based method the analysis of molecular structures, a method of assessing the ability of the potential modulator to interact in the binding site on the PFK surface, a method of providing data for generating structures and/or performing design for ligands binding PFK, PFK homologues or analogues, complexes of PFK with a potential modulator, or complexes of PFK homologues or analogues with potential modulators, a computer system. The invention makes it possible to design a suitable activator, which could be used to modulate PFK activity, taking advantage of PFKs capability to bind in its effector site phosphate groups or other groups with a similar potential to interact, which correspond with the substituents in positions 1, 2 and 6 of the fructose ring, or groups corresponding with positions of the fructose ring groups interacting in the PFK effector site. 
     Glycolysis is the basis of anaerobic and aerobic metabolism processes and occurs in almost all organisms (Fothergill-Gilmore &amp; Michels, 1993). It is the main energy source in many prokaryotes and in the eukaryotic cell types devoid of mitochondria or functioning under low oxygen or anaerobic conditions. During glycolysis one glucose molecule is converted to two molecules of pyruvate while two molecules of ATP are produced. The rate of glycolysis is tightly regulated depending on the cell&#39;s needs for energy and building blocks for biosynthetic reactions. 
     Phosphofructokinase (PFK, EC 2.7.1.11) is the main control point in the glycolytic pathway. PFK catalyses the conversion of fructose-6-phosphate (Fru-6-P) to fructose-1,6-diphosphate (Fru-1,6-P 2 ) with the simultaneous conversion of ATP to ADP. The reaction releases much of energy, therefore it is practically irreversible. The reverse reaction is catalysed by a fructose-1,6 bisphosphatase (Benkovic &amp; de Maine, 1982). PFK is the enzyme with the most complex regulatory mechanism in the glycolytic pathway. The major isozyme of PFK is PFK1, a multi-subunit oligomeric allosteric enzyme whose activity is modulated by a number of effectors. In general, PFK is sensitive to the “energy level” of the cell, as indicated by the levels of ATP relative to the products of ATP hydrolysis, but the mechanisms of control are different in eukaryotes and prokaryotes. In the relatively well studied prokaryotes PFK is activated in response to “low energy level”, i.e. by the products of ATP hydrolysis, while in eukaryotes PFK is inhibited by ATP (“high energy”) and activated by AMP and, to a lesser extent, by ADP (Hofmann &amp; Kopperschläger, 1982). In a classic case of feedback inhibition PFKs are also inhibited by citrate, allowing feedback from the citric acid cycle (Sols, 1981). In eukaryotes, but not in prokaryotes, PFKs are also activated by fructose-2,6-diphosphate (Fru-2,6-P 2 ). This potent allosteric regulator, which also inhibits the corresponding bisphosphatase, reflects a higher level of complexity of eukaryotes, compared to prokaryotes. It overrides the inhibition by ATP, which is essential in some tissues (e.g. in muscles), and makes PFK sensitive to the action of the hormones: glucagon and insulin in “higher” organisms (Pilkis et al., 1988; Okar &amp; Lange, 1999). 
     PFK1 is an allosteric enzyme, showing a characteristic sigmoidal activity profile instead of the common Michaelis-Menten kinetics. The sigmoidal profile indicates co-operativity between the active sites in the oligomeric enzyme. At low substrate concentrations the enzyme shows little activity resulting from its low affinity for the substrate. As the concentration of the substrate increases, so does the substrate affinity and the enzymatic activity. This behaviour is explained in terms of a balance between two alternative forms of the enzyme: the inactive “T-state” and the active “R-state”. The ground state of the enzyme is the T-state. It predominates in the absence of the allosteric substrates and allosteric activators. These ligands bind only to the R-state. The binding stabilises the active form, thus enabling more substrate molecules to bind to the still vacant binding sites in the oligomeric enzyme. In the overall effect, the allosteric substrate and the allosteric activators shift the balance in solution from the enzyme&#39;s inactive ground T-state towards the active R-state. 
     In the past, crystallographic studies focused on PFK1 from prokaryotes:  E. coli  and  B. stearothermophilus . Prokaryotic PFK1 is a homotetramer with a subunit molecular weight of approximately 37 kDa. In the 1980s, the control mechanism of bacterial PFK1 was investigated by means of protein crystallography. The T- and the R-states of the  E. coli  and  B. stearothermophilus  enzymes were explained in terms of the proteins&#39; quaternary structure. The transitions between the T- and R-states involves a rearrangement of the enzymes&#39; subunits. The binding sites of the substrate, Fru-6-P, and the allosteric effectors span the inter-subunit interface of the R-state. In the T-state, these ligands cannot bind (Evans &amp; Hudson, 1979; Evans et al., 1981; Shirakihara &amp; Evans 1988; Rypniewski &amp; Evans, 1989; Schirmer &amp; Evans, 1990). 
     The structures of eukaryotic PFK1 are more complex than in prokaryotes. So is their control mechanism. In yeast, PFK1 is an oligomer of the form α 4 β 4 . Each subunit is more than twice the size of the prokaryotic PFK1 subunit. The amino acid sequence of each eukaryotic subunit consists of two homologous parts, each half being homologous to a prokaryotic PFK1 subunit. The two types of subunits, α and β, are also homologous in sequence. It has been postulated that yeast PFK1 is the result of two gene duplications: the first tandem duplication resulted in a subunit of double size compared to the prokaryotic subunit, and the second gene duplication created two subunit types (Poorman et al., 1984; Heinisch et al, 1989). 
     Attempts were made to extend the crystallographic study to include PFK1 in eukaryotes but no suitable crystals could be obtained. Much data were obtained on the eukaryotic PFK by means of biochemistry, enzyme kinetics, genetics, mutagenesis and single-particle electron microscopy but detailed structural information was unavailable. 
     In the case of PFK1, the allosteric substrate is Fru-6-P (the other substrate, ATP, has no allosteric effect) and there are several allosteric activators of which the most potent is Fru-2,6-P 2 , found only in eukaryotes. It abolishes the inhibitory effect of ATP. The Fru-2,6-P 2  binding site, which is part of the regulatory mechanism in eukaryotes, had been proposed, based on amino the acid sequence analysis, which evolved from the active sites that became redundant after the gene duplication and, losing the catalytic function, acquired a regulatory role (Heinisch et al. 1996). However, the Fru-2,6-P 2  binding site has not been described until now in terms of a three-dimensional atomic model. 
     In the patent application MXPA05011769 (publ. 2006-01-26) crystal of PDE5, its crystalline structure and its use in drug design were presented. The invention relates to the soakable crystals of a phosphodiesterase 5 (PDE5) and their uses in identifying PDE5 ligands, including PDE5 ligands and inhibitor compounds. The present invention also relates to methods of identifying such PDE5 inhibitor compounds and their medical use. The present invention additionally relates to crystals of PDE5 into which ligands may be soaked and crystals of PDE5 10 comprising PDE5 ligands that have been soaked into the crystal. 
     In the patent application EP 0130030 (publ. 1985-01-02) diagnostic application of phosphofructokinase was described. In the patent application WO2005083069 (publ. 2005-09-09) PDE2 crystal structures for structure based drug design were described. Crystalline compositions of Phosphodiesterase Type 2 (PDE2), particularly of the PDE2 catalytic domain, and amino acid sequences utilized to form such crystalline compositions, which are used to screen PDE2 ligands. The ligands are formulated into pharmaceutical compositions, and used for treatment of disease states or disorders mediated by PDE2. In the patent application WO 2005083069 (publ. 2005-09-09) PDE2 crystal structures for structure based drug design was described. A crystalline compositions of Phosphodiesterase Type 2 (PDE2), particularly of the PDE2 catalytic domain, and amino acid sequences utilized to form such crystalline compositions, which are used to screen PDE2 ligands. The ligands are formulated into pharmaceutical compositions, and used for treatment of disease states or disorders mediated by PDE2. 
     In the patent application US2006110743 (publ. 2006-05-25) drug evolution: drug design at “hot spots” was described. A new method of designing and generating compounds having an increased probability of being drugs, drug candidates, or biologically active compounds, in particular having a therapeutic utility, is disclosed. The method consists of identifying a group of bioactive compounds, preferably of diverse therapeutic uses or biological activities and built on a common building block. In this group of compounds, side chains modifying the building block are identified and used to generate a second set of compounds according to the proposed methods of “hybridization”, “single substitution” or “incorporation of frequently used side chains”. If the compounds in the second set built on the same building block contain an unusually large number of drugs, preferably with diverse therapeutic uses or biological activities, they constitute a “hot spot”. A focused combinatorial library of the “hot spot” is then generated, preferably by methods of combinatorial chemistry, and compounds of this library are screened for a variety of therapeutic uses or biological activities. The method generates drugs, drug candidates, or biologically active compounds with a high probability, without requiring any prior knowledge of biological targets. 
     Despite the above-described compounds and methods of designing and generating drugs, drug candidates or biologically active compounds, methods for identifying modulators for metabolic pathways, comprising screening for agents that modulate the activity of enzymes, computer-assisted methods of structure based drug design of different inhibitors using a 3-D structure of proteins, still there is a need for a successful design of an efficient activator or inhibitor which could be used to modulate the PFK activity and designed to exploit the PFK&#39;s possibilities to bind at its effector site the phosphate or other groups with similar binding potential corresponding to the ligands at positions 1, 2 and 6 of the fructose ring or groups corresponding with positions of groups of fructose ring, which interact with the effector binding site of PFK. 
     The goal of the present invention is to provide a method which may be used to obtain stable compounds which would act on PFK analogously to Fru-2,6-P 2 , and which could be used as activators of this enzyme. 
     The fulfillment of this goal and the solution of problems regarding the design of compounds which can interact with the Fru-2,6-P 2  effector binding site and induce enzymatic activity of PFK, as well as the development of the atomic model which enables the design of artificial activators, have been achieved in the present invention. 
     The subject of invention is a crystallographic model of the binding site wherein it is a part of the eukaryotic phosphofructokinase (PFK), in complex with the allosteric activator D-fructose-2,6-bisphosphate (Fru-2,6-P 2 ), wherein the atomic coordinates x, y, z of a portion of PFK which determine two homologous binding sites of the activator (effector), including the bound Fru-2,6-P 2  molecules, are presented in Tables 1a and 1b, or a derivative set of transformed coordinates expressed in any reference system. 
     Preferably, the amino acid residues from Tables 1a or 1b are substituted with the amino acid residues present in a homologous sequence of other eukaryotic PFK.
 
Preferably, the three-dimensional structure described by means of the atomic coordinates x, y, z, after being superimposed with the least squares minimization method, having the root mean square deviation equal or less than 0.1 nm, in relation to the atomic coordinates x, y, z presented in Tables 1a and 1b.
 
     The next subject of the invention is a modulator which regulates the catalytic activity of PFK, wherein said modulator is a compound presented on  FIG. 1 , where A and C are selected from among the groups: —PO 4 , —SO 4  or —C—SO 2 O - , B is one of the bridges: —O— or —S—; D is selected from among the groups —PO 4 , —SO 4 , —OH or —C—SO 2 O - , E is —H, # is an atom C with hybridization sp 3 ; R1 and R2 are either —CXH—OH or —CX═O or —H, where X is a hydrogen atom or bonds with other R groups or bonds with other R groups through the —CH 2 —, group; and the —CH 2 — groups are between D and # and between C and #. Preferably, the modulator stimulates the catalytic activity of PFK. 
     The next subject of invention is a method of designing a PFK modulator, wherein the modulator is a compound of the formula presented in  FIG. 1 , and where A and C are selected from among the groups: —PO 4 , —SO 4  or —C—SO 2 O - ; B is one of the bridges —O— or —S—; D is selected from among the groups —PO 4 , —SO 4 , —OH or —C—SO 2 O - ; E is —H, # is an atom C with hybridization sp 3 ; R1 and R2 are either —CXH—OH or —CX═O or —H, where X is a hydrogen atom or bonds with other R groups or bonds with other R groups through the —CH 2 —, group; and the —CH 2 — groups are between D and # and between C and #. 
     Preferably, the modulator design includes:
         a) exploring the PFK atomic coordinates which constitute the binding site of the PKF effector presented in tables 1a and 1b to obtain information about the three-dimensional structure and electrostatic properties of the protein surface;   b) designing a PFK modulator using the effector binding site information given in Tables 1a or 1b.       

     The next subject of the invention is a method of selecting the PFK modulator, wherein the modulator is a compound of the formula presented in  FIG. 1 , and where A and C are selected from among the groups: —PO 4 , —SO 4  or —C—SO 2 O - , B is one of the bridges: —O— or —S—; D is selected from among the groups —PO 4 , —SO 4 , —OH or —C—SO 2 O - ; E is —H, # is an atom C with hybridization sp 3 , R1 and R2 are either —CXH—OH or —CX═O or —H, where X is a hydrogen atom or bonds with other R groups or bonds with other R groups through the —CH 2 —, group; and the —CH 2 — groups are between D and # and between C and #. 
     Preferably the modulator design includes:
         a) exploring the PFK atomic coordinates which constitute the binding site of the PKF effector, presented in Tables 1a and 1b to obtain information about the structure and properties of the protein surface;   b) selecting a PFK modulator using the effector binding site information given in tables 1a or 1b.       

     The next subject of invention is a method of producing the PFK modulator, characterised in that it comprised the identification of a compound or designing a compound that fits into the effector site binding pocket of PFK in its uninhibited conformation, wherein the said conformation of the effector site binding pocket of PFK is defined by the x, y, z-coordinates of atoms in the set of amino acid residues given in Tables 1a or 1b. 
     The next subject of invention is a computer-based method for the analysis of the interaction of a molecular structure with PFK, characterised in that it comprises:
         a) providing a structure comprising a three-dimensional representation of PFK effector binding site, whose representation comprises all or a portion of the coordinates given in Tables 1a or 1b, or coordinates whose differences from those are within a root-mean-square deviation (r.m.s.d.) equal or less than 0.1 nm,   b) providing a molecular structure to be fitted to the said PFK effector binding site; and   c) fitting the molecular structure to the PFK structure of a).       

     The next subject of the invention is a computer-based method for the analysis of molecular structures which comprises:
         a) providing the coordinates of at least two atoms of a PFK structure as defined in Tables 1a or 1b, wherein the root mean square deviation for the atoms is equal or less than 0.1 nm (“selected coordinates”);   b) providing a molecular structure to be fitted to the selected coordinates.       

     The next subject of invention is a method of assessing the ability of a candidate modulator to interact in the binding site on the PFK surface, characterised in that it comprises the steps of:
         a) obtaining or synthesising the said candidate modulator;   b) forming a crystallized complex of a PFK protein whose atomic coordinates x, y, z include the coordinates presented in Tables 1a or 1b, or the set of coordinates of a homologous part of protein or candidate modulator expressed in any reference system which, after being superimposed with the least squares minimization method, has the root mean square deviation equal or less than 0.1 nm, in relation to the atomic coordinates x, y, z presented in Tables 1a and 1b;   c) analysing the said complex by X-ray crystallography or NMR spectroscopy to determine the ability of the said candidate modulator to interact with the binding site on the PFK surface.       

     The next subject of invention is a method of providing data for generating structures and/or designing the drugs which bind the PFK, PFK homologues or analogues, complexes of PFK, or complexes of the PFK homologues or analogues with potential modulators, wherein communication is established with a remote device which contains the computer-readable data comprising at least one of:
         a) atomic coordinate data presented in Tables 1a or 1b, or a set of coordinates expressed in any reference system which, after being superimposed with the least squares minimization method, has the root mean square deviation equal or less than 0.1 nm, when such data defines the three-dimensional structure of the effector-binding site on the PFK surface;   b) atomic coordinate data of a target effector binding site on the PFK homologue or analogue surface, generated by homology modelling of the target based on the data from Tables 1a or 1b, with the root mean square deviation from atoms equal or less than 0.1 nm;   c) receiving the said computer-readable data from a remote device.       

     The next subject of invention is a computer system, characterized in that it contains at least one of:
         a) atomic coordinate data presented in Tables 1a or 1b, or such data which, after being superimposed with the least squares minimization method, has the root mean square deviation from the atoms in Tables 1a or 1b equal or less than 0.1 nm, where such data defines the three-dimensional structure of the effector-binding site on the PFK surface or at least its selected coordinates;   b) atomic coordinate data of an effector binding site on the surface of target PFK protein, generated by homology modelling of the target based on the data from Tables 1a or 1b, when the root mean square deviation from atoms from Tables 1a or 1b, after being superimposed with the least squares minimization method, is equal or less than 0.1 nm;   c) atomic coordinate data of an effector binding site on the surface of the target PFK protein, generated by the interpretation of data obtained from the analysis of the X-ray crystallography or NMR, based on the data from Tables 1a or 1b, when the root mean square deviation of atoms from Tables 1a or 1b, after being superimposed with the least squares minimization method, is equal or less than 0.1 nm, and/or   d) crystallographic structure factors, obtained from the atomic coordinates (c) or (d).       

    
    
     
       The attached figures facilitate a better understanding of the nature of the present invention. 
         FIG. 1  presents a scheme of a modulator where A and C are selected from —PO 4 , —SO 4  or —C—SO 2 O -  groups, B is selected from —O— or —S— bridge, D is selected from —PO 4 , —SO 4 , —OH or —C—SO 2 O -  group, E is —H, # is atom C with hybridization sp 3 , R1 and R2 are selected from —CXH—OH or —CX═O or —H, where X is a hydrogen atom or a bond to the other R-group or a bond to the other R-group via a —CH2- group. Especially important for the modulator is group C. In case of the activator C is sulphate, phosphate or sulphonic group and interacts with the neighboring subunit of PFK stabilizing the enzyme in the active R-state. 
       Table 1a and 1b show atomic coordinates in PDB (Protein Data Bank) format of parts of crystallographic atomic PFK from  S. cerevisiae  (yeast), which are the effector sites on the protein surface, and associated molecules of the effector Fru-2,6-P 2 . Table 1a shows the location of the α-chain on the surface and Table 1b shows the appropriate site for the β-chain. These coordinates are empirically defined as a result of a crystallographic analysis and are a starting point for designing the modulator of the enzymatic activity of PFK whose characteristics were described in  FIG. 1 . 
         FIG. 2  and  FIG. 3  show a representative structures of the effector-binding site in α chain ( FIG. 2 ) and β chain ( FIG. 3 ) These chains in the PFK enzyme are similar but not identical. The PFK molecule is a heterooctamer α 4 β 4 . Bound effector molecules, fructose-2,6-bisphosphate (FDP-5, FDP-2) are also depicted. 
         FIG. 4  and  FIG. 5  are similar to  FIGS. 2 and 3  but they additionally show a rendering of the molecular surface to illustrate the shape of the effector binding cavity. 
         FIG. 6  is a scheme of interactions within the effector binding site, between the PFK and the fructose-2,6-bisphosphate effector. Amino acid residues from the α chain are indicated on a white background while residues from the β chain are shown on a grey background. 
     
    
    
     Below, there are example embodiments of the present invention defined above. 
     Determination of the Crystalline Structure of Phosphofructokinase (PFK) in Complex with fructose-6-phosphate (F-6-P) and fructose-2,6-bis-phosphate (Fru-2,6-P2) 
     The PFK from baker&#39;s yeast ( Saccharomyces cerevisiae ) was prepared according to the Hoffmann and Kopperschlaeger method (1982). The native form of the enzyme, called 21S, was subjected to a limited proteolysis with alpha-chymotrypsin at a ratio of 1:600 to PFK, in the presence of 5 mM ATP. The tetrameric form 12S created as a result of the digestion was subsequently precipitated with ammonium sulfate, the centrifuged sediment was dissolved and dialysed, and then passed twice through the Resource Q column in the HPLC system to remove ATP and low molecular weight proteolytic forms. Protein purified this way was dialysed at 277 K in 20 mM of HEPES, pH 7.4, 1 mM EDTA, 0.2 M sodium acetate, 0.1 M ammonium sulfate, 2 mM DTT, 0.1 mM PMSF and 10 mM F-6-P. The crystals were grown by vapour diffusion in hanging drops at 277 K with a reservoir solution containing 6-10% PEG4000, 0.2 mM sodium acetate in 0.1 M MES buffer, pH 6.0. The crystallization drop initially consisted of 3 microlitres of protein at the mg/ml concentration 8 and 3 microlitres of reservoir solution. Crystals, in the form of long needles of the 0.2 mm diameter, appeared within about two weeks. 
     The crystallographic data was recorded using synchrotron radiation from the crystal under cryo-conditions at 100 K and stabilized with a solution which comprised glycerol in the 20% concentration (by volume), the components of the reservoir solution, and the ligands Fru-6-P and Fru-2,6-P 2 . Crystallographic data was processed with the HKL software package (Otwinowski and Minor, 1997). The crystals belonged to the spatial group P2(1)2(1)2(1) and the dimensions of a unit cell were a=18.0 nm, b=18.6 nm, c=23.7 nm. The crystal structure was determined by molecular replacement (MR) using the PFK tetramer from  E. coli  as the search model (Shirakihara et al., 1988, PDB code 1pfk). In the determination of the structure important was also the information on the shape of the molecule obtained from electron microscopy (Ruiz et al., 2001). The MR calculations were performed with the AMoRe software (Navaza, 1994). The model obtained via MR method consisted initially of four tetrameric molecules of a bacterial PFK. This model was used to calculate the phases. These phases with experimentally determined amplitudes of X-ray scattering structure factors were used to calculate an electron density map. The map was subjected to non-crystallographic symmetry averaging and solvent flattening with the DM software (Cowtan, 1994). The resulting map significantly differed from the initial one which had systematic errors caused by the inaccurate initial model. The phases and the resolution were extended in the course of numerous DM cycles, from the initial values of 0.15-0.04 nm to the final range of 0.35-0.29. The proof of correctness of the performed phase refinement was the fact that the obtained electron density map included details which were absent in the initial model, for instance the ligand molecules Fru-6-P and Fru-2,6-P 2 , and amino acid residues absent in the PFK molecule from  E. coli . Subsequently, the chains with target eukaryotic sequence ( Saccharomyces cerevisiae ) were built into the electron density map, and the atomic model was refined with the CNS program (Brunger et al., 1998). The refinement cycles were interspersed with manual model corrections based on the maps 2Fo-Fc and Fo-Fc. The temperature factors were not refined initially, but with time they were refined for individual side chains and separately for the main chain atoms of individual amino acid residues. The statistics, such as R-factor and R-free (based on 5% of reflections) were monitored, as well as the FOM (figure of merit) (Brunger, 1992). The final values of those statistics were as follows: R=0.238, R-free=0.311, FOM=0.73. The atomic model was validated with the PROCHECK software (Laskowski et al., 1993). All indices were within tolerance limits, or better than what could be expected for a structure determined at such a resolution. For instance, on the Ramachandran plot 79.0% of the amino acid residues are located in the most favoured region, whereas the expected value for the 0.029 nm resolution is 68.7%. The 2Fo-Fc electron density map allows an unambiguous determination of the course of the polypeptide chains and the correct “register” of sequences, because both the density for the main chain and the typical shapes of side chains made such a determination possible. All four independent models of α chains and the β chains are consistent with each other. The parts of the molecule which have their equivalents in the sequence of PFK from  E. coli  are also consistent in terms of the spatial structure. 
     The final model includes more than six thousand amino acid residues which constitute eight polypeptide chains (four α and four β), eight molecules of Fru-6-P and eight molecules of Fru-2,6-P 2 . These chains, situated in the asymmetric unit of the crystallographic unit cell, have a form very similar to the general shape of the molecule as it was determined by electron microscopy (EM), despite the fact that the crystal contains the 12S molecules, that is the form obtained as a result of partial proteolysis (see above) and being tetrameric in solution, while the EM results are for the native octameric 21 S form. It is however evident that in the crystalline structure the tetrameric 12S molecules associate in pairs, just like in the native form, and create an octamer in which four α subunits constitute the core of the molecule and the β subunits are on the outside. The structures of α and β chains are similar (just like the amino acid sequences), each of them resembling a dimer of the PFK subunits from  E. coli . The α and β chains associate in pairs so that their dimer structure resembles the tetramers of the PFK subunits from the bacteria. Four such dimers associate and create the octameric PFK molecule from  S. cerevisiae . The determined structure corresponds to the active form (R-state) that is the quaternary form of the molecule, which allows the binding of the Fru-6-P substrate. The binding of Fru-2,6-P 2  in the effector site is analogous to the binding of Fru-6-P in the substrate binding site. Both these ligands are visible in the crystal structure. The other argument for the R form in this crystalline structure is also the comparisons with the structures of bacterial PFK of both forms. The interactions between the subunits in the eukaryote structure are comparable to the active structures of the PFK bacterial forms (R-state), and not to inactive forms (T-state). 
     The crystalline structure of PFK from  S. cerevisiae  is the first structure of eukaryotic PFK regulated by Fru-2,6-P 2  (which is typical for eukaryotes) to have been determined experimentally in terms of a three-dimensional atomic model. The model shows detailed interactions of PFK with the activator Fru-2,6-P 2  and explains the mechanism of its action. The effector binds between two subunits and stabilizes the quaternary structure of the enzyme in the active form (R-state). Such a role of the Fru-2,6-P 2  enzyme was earlier postulated on the basis of the evolution consideration, but only here the presented atomic structure of the effector binding site along with the associated Fru-2,6-P 2  molecule allows an understanding of this mechanism in terms of actual interatomic interactions, such as hydrogen bonds or the Van der Waals forces. The accurate determination of the ligand bond site allows also determination of an optimum match of the ligand molecule in the steric sense. The presented model of the effector-ligand binding site is the key to control the activity of the eukaryotic PFK ( FIGS. 1 to 6 ). 
     The detailed model of the effector binding site of PFK and its interactions with Fru-2,6-P 2  constitute a matrix for the structure-based design of compounds different than the native ligand Fru-2,6-P 2  but sharing with it some common features; the compounds are strong activators or inhibitors of this PFK or related enzymes. It is possible that Fru-1,6-P 2  and Fru-6-P can also bind at the effector binding site. The analysis of the atomic model of the PFK in complex with Fru-2,6-P 2  indicates such a possibility. The PFK surface has a suitable cavity which could accommodate the phosphate group bonded with 1-carbon of the fructose ring. The designing of an artificial effector involves taking advantage of the possibility of fitting and specifically binding appropriate groups in the cavity that constitutes the Fru-2,6-P 2  binding site on the surface of the eukaryotic PFK. The proposed compounds are designed in analogy to the molecule Fru-2,6-P 2  bound in the effector positions on the PFK surface (Tables 1a and 1b). Such a compound has the ability to bind PFK in the sites corresponding to the phosphate group bound with the atom C6 of the fructose ring, and binding PFK in the position corresponding to the phosphate group bound at the atom C2, or in the position corresponding to the substituent at the atom C1, or in the positions corresponding to the fructose ring groups which are capable of making hydrogen bonds. 
       FIG. 6  provides a summary of the experimentally determined interactions in the effector site. The analysis of such interactions, with due consideration of steric conditions, has resulted in determining the “active part”, or a pharmacofore, of an artificial ligand which would have activator properties and also one which would be a PFK inhibitor. The “active part” is presented in  FIG. 1 . The activator should have as many as possible of the features defined in the presented diagram and description to  FIG. 1 . The most important for the activator is the presence of the “C” group corresponding to 6-phosphate in the natural activator Fru-2,6-P 2 . Whereas the larger part of the ligand is bound to one PFK subunit, the “C” group binds a neighbouring subunit and stabilizes the quaternary structure of the enzyme in the active form (R-state). 
     An obvious example of using this invention is the activation of yeast metabolism in industrial applications, such as the brewing industry, distilling industry, and industrial chemical synthesis. The yeasts have a limited tolerance to ethanol which they produce. At a given alcohol concentration, the yeast cells cease to be active. The yeast tolerance limits is well defined, but hard to overcome. If it could be extended even by a small amount, the fermentation results on industrial scale would improve significantly. Using the artificial activator in the fermentation process, particularly in its final stage, should stimulate higher activity of the yeast cells, against their natural tendency to limit the activity level when the alcohol concentration reaches a critical level. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 1a 
               
               
                   
               
             
            
               
                 ATOM 
                 33233 
                 N 
                 ARG 
                 F 
                 754 
                 −8.057 
                 −8.485 
                 121.415 
                 1.00 
                 60.96 
                 F 
                 N 
               
               
                 ATOM 
                 33234 
                 CA 
                 ARG 
                 F 
                 754 
                 −7.594 
                 −9.038 
                 122.682 
                 1.00 
                 61.68 
                 F 
                 C 
               
               
                 ATOM 
                 33235 
                 CB 
                 ARG 
                 F 
                 754 
                 −7.961 
                 −8.121 
                 123.844 
                 1.00 
                 51.97 
                 F 
                 C 
               
               
                 ATOM 
                 33236 
                 CG 
                 ARG 
                 F 
                 754 
                 −7.836 
                 −8.789 
                 125.217 
                 1.00 
                 49.71 
                 F 
                 C 
               
               
                 ATOM 
                 33237 
                 CD 
                 ARG 
                 F 
                 754 
                 −7.972 
                 −7.763 
                 126.304 
                 1.00 
                 49.64 
                 F 
                 C 
               
               
                 ATOM 
                 33238 
                 NE 
                 ARG 
                 F 
                 754 
                 −6.931 
                 −6.750 
                 126.153 
                 1.00 
                 49.96 
                 F 
                 N 
               
               
                 ATOM 
                 33239 
                 CZ 
                 ARG 
                 F 
                 754 
                 −6.783 
                 −5.698 
                 126.953 
                 1.00 
                 49.95 
                 F 
                 C 
               
               
                 ATOM 
                 33240 
                 NH1 
                 ARG 
                 F 
                 754 
                 −7.616 
                 −5.511 
                 127.971 
                 1.00 
                 49.14 
                 F 
                 N 
               
               
                 ATOM 
                 33241 
                 NH2 
                 ARG 
                 F 
                 754 
                 −5.794 
                 −4.839 
                 126.738 
                 1.00 
                 48.11 
                 F 
                 N 
               
               
                 ATOM 
                 33242 
                 C 
                 ARG 
                 F 
                 754 
                 −8.076 
                 −10.433 
                 123.025 
                 1.00 
                 62.07 
                 F 
                 C 
               
               
                 ATOM 
                 33243 
                 O 
                 ARG 
                 F 
                 754 
                 −9.244 
                 −10.781 
                 122.820 
                 1.00 
                 63.60 
                 F 
                 O 
               
               
                 ATOM 
                 33926 
                 N 
                 LYS 
                 F 
                 847 
                 −4.159 
                 −10.783 
                 128.058 
                 1.00 
                 51.37 
                 F 
                 N 
               
               
                 ATOM 
                 33927 
                 CA 
                 LYS 
                 F 
                 847 
                 −4.101 
                 −10.402 
                 129.456 
                 1.00 
                 52.31 
                 F 
                 C 
               
               
                 ATOM 
                 33928 
                 CB 
                 LYS 
                 F 
                 847 
                 −5.087 
                 −9.266 
                 129.772 
                 1.00 
                 49.97 
                 F 
                 C 
               
               
                 ATOM 
                 33929 
                 CG 
                 LYS 
                 F 
                 847 
                 −4.612 
                 −7.861 
                 129.409 
                 1.00 
                 50.20 
                 F 
                 C 
               
               
                 ATOM 
                 33930 
                 CD 
                 LYS 
                 F 
                 847 
                 −3.440 
                 −7.402 
                 130.273 
                 1.00 
                 50.36 
                 F 
                 C 
               
               
                 ATOM 
                 33931 
                 CE 
                 LYS 
                 F 
                 847 
                 −3.092 
                 −5.948 
                 130.013 
                 1.00 
                 48.67 
                 F 
                 C 
               
               
                 ATOM 
                 33932 
                 NZ 
                 LYS 
                 F 
                 847 
                 −4.214 
                 −5.059 
                 130.402 
                 1.00 
                 48.72 
                 F 
                 N 
               
               
                 ATOM 
                 33933 
                 C 
                 LYS 
                 F 
                 847 
                 −4.461 
                 −11.612 
                 130.288 
                 1.00 
                 52.57 
                 F 
                 C 
               
               
                 ATOM 
                 33934 
                 O 
                 LYS 
                 F 
                 847 
                 −5.297 
                 −12.432 
                 129.898 
                 1.00 
                 53.90 
                 F 
                 O 
               
               
                 ATOM 
                 26243 
                 N 
                 ALA 
                 E 
                 603 
                 −11.442 
                 1.471 
                 129.846 
                 1.00 
                 46.04 
                 E 
                 N 
               
               
                 ATOM 
                 26244 
                 CA 
                 ALA 
                 E 
                 603 
                 −11.603 
                 0.034 
                 129.897 
                 1.00 
                 45.47 
                 E 
                 C 
               
               
                 ATOM 
                 26245 
                 CB 
                 ALA 
                 E 
                 603 
                 −10.989 
                 −0.533 
                 131.163 
                 1.00 
                 71.43 
                 E 
                 C 
               
               
                 ATOM 
                 26246 
                 C 
                 ALA 
                 E 
                 603 
                 −13.113 
                 −0.172 
                 129.894 
                 1.00 
                 44.51 
                 E 
                 C 
               
               
                 ATOM 
                 26247 
                 O 
                 ALA 
                 E 
                 603 
                 −13.884 
                 0.734 
                 130.254 
                 1.00 
                 43.55 
                 E 
                 O 
               
               
                 ATOM 
                 26706 
                 N 
                 ARG 
                 E 
                 665 
                 −8.389 
                 2.568 
                 125.763 
                 1.00 
                 45.24 
                 E 
                 N 
               
               
                 ATOM 
                 26707 
                 CA 
                 ARG 
                 E 
                 665 
                 −7.739 
                 3.675 
                 126.471 
                 1.00 
                 45.83 
                 E 
                 C 
               
               
                 ATOM 
                 26708 
                 CB 
                 ARG 
                 E 
                 665 
                 −6.692 
                 3.124 
                 127.446 
                 1.00 
                 46.97 
                 E 
                 C 
               
               
                 ATOM 
                 26709 
                 CG 
                 ARG 
                 E 
                 665 
                 −7.242 
                 2.268 
                 128.567 
                 1.00 
                 46.99 
                 E 
                 C 
               
               
                 ATOM 
                 26710 
                 CD 
                 ARG 
                 E 
                 665 
                 −6.099 
                 1.757 
                 129.445 
                 1.00 
                 48.49 
                 E 
                 C 
               
               
                 ATOM 
                 26711 
                 NE 
                 ARG 
                 E 
                 665 
                 −6.571 
                 0.819 
                 130.462 
                 1.00 
                 49.00 
                 E 
                 N 
               
               
                 ATOM 
                 26712 
                 CZ 
                 ARG 
                 E 
                 665 
                 −7.375 
                 1.143 
                 131.472 
                 1.00 
                 49.18 
                 E 
                 C 
               
               
                 ATOM 
                 26713 
                 NH1 
                 ARG 
                 E 
                 665 
                 −7.804 
                 2.386 
                 131.628 
                 1.00 
                 50.34 
                 E 
                 N 
               
               
                 ATOM 
                 26714 
                 NH2 
                 ARG 
                 E 
                 665 
                 −7.772 
                 0.219 
                 132.326 
                 1.00 
                 50.65 
                 E 
                 N 
               
               
                 ATOM 
                 26715 
                 C 
                 ARG 
                 E 
                 665 
                 −7.095 
                 4.757 
                 125.602 
                 1.00 
                 44.81 
                 E 
                 C 
               
               
                 ATOM 
                 26716 
                 O 
                 ARG 
                 E 
                 665 
                 −6.649 
                 5.779 
                 126.122 
                 1.00 
                 44.15 
                 E 
                 O 
               
               
                 ATOM 
                 26931 
                 N 
                 GLU 
                 E 
                 694 
                 −9.377 
                 8.324 
                 131.199 
                 1.00 
                 40.89 
                 E 
                 N 
               
               
                 ATOM 
                 26932 
                 CA 
                 GLU 
                 E 
                 694 
                 −8.965 
                 8.635 
                 129.831 
                 1.00 
                 40.87 
                 E 
                 C 
               
               
                 ATOM 
                 26933 
                 CB 
                 GLU 
                 E 
                 694 
                 −8.605 
                 7.373 
                 129.052 
                 1.00 
                 57.96 
                 E 
                 C 
               
               
                 ATOM 
                 26934 
                 CG 
                 GLU 
                 E 
                 694 
                 −7.149 
                 6.976 
                 129.157 
                 1.00 
                 58.96 
                 E 
                 C 
               
               
                 ATOM 
                 26935 
                 CD 
                 GLU 
                 E 
                 694 
                 −6.761 
                 6.489 
                 130.542 
                 1.00 
                 59.23 
                 E 
                 C 
               
               
                 ATOM 
                 26936 
                 OE1 
                 GLU 
                 E 
                 694 
                 −7.193 
                 5.377 
                 130.927 
                 1.00 
                 57.97 
                 E 
                 O 
               
               
                 ATOM 
                 26937 
                 OE2 
                 GLU 
                 E 
                 694 
                 −6.027 
                 7.229 
                 131.236 
                 1.00 
                 59.39 
                 E 
                 O 
               
               
                 ATOM 
                 26938 
                 C 
                 GLU 
                 E 
                 694 
                 −10.072 
                 9.368 
                 129.091 
                 1.00 
                 42.65 
                 E 
                 C 
               
               
                 ATOM 
                 26939 
                 O 
                 GLU 
                 E 
                 694 
                 −9.802 
                 10.148 
                 128.176 
                 1.00 
                 43.60 
                 E 
                 O 
               
               
                 ATOM 
                 27157 
                 N 
                 THR 
                 E 
                 722 
                 −16.743 
                 3.679 
                 134.668 
                 1.00 
                 47.10 
                 E 
                 N 
               
               
                 ATOM 
                 27158 
                 CA 
                 THR 
                 E 
                 722 
                 −15.591 
                 2.853 
                 135.000 
                 1.00 
                 47.36 
                 E 
                 C 
               
               
                 ATOM 
                 27159 
                 CB 
                 THR 
                 E 
                 722 
                 −14.299 
                 3.700 
                 135.021 
                 1.00 
                 34.01 
                 E 
                 C 
               
               
                 ATOM 
                 27160 
                 OG1 
                 THR 
                 E 
                 722 
                 −13.200 
                 2.895 
                 135.455 
                 1.00 
                 34.33 
                 E 
                 O 
               
               
                 ATOM 
                 27161 
                 CG2 
                 THR 
                 E 
                 722 
                 −14.461 
                 4.896 
                 135.965 
                 1.00 
                 32.86 
                 E 
                 C 
               
               
                 ATOM 
                 27162 
                 C 
                 THR 
                 E 
                 722 
                 −15.852 
                 2.311 
                 136.390 
                 1.00 
                 47.81 
                 E 
                 C 
               
               
                 ATOM 
                 27163 
                 O 
                 THR 
                 E 
                 722 
                 −16.289 
                 3.042 
                 137.264 
                 1.00 
                 47.67 
                 E 
                 O 
               
               
                 ATOM 
                 27164 
                 N 
                 VAL 
                 E 
                 723 
                 −15.593 
                 1.028 
                 136.587 
                 1.00 
                 48.94 
                 E 
                 N 
               
               
                 ATOM 
                 27165 
                 CA 
                 VAL 
                 E 
                 723 
                 −15.797 
                 0.399 
                 137.886 
                 1.00 
                 50.18 
                 E 
                 C 
               
               
                 ATOM 
                 27166 
                 CB 
                 VAL 
                 E 
                 723 
                 −15.318 
                 −1.069 
                 137.875 
                 1.00 
                 29.18 
                 E 
                 C 
               
               
                 ATOM 
                 27167 
                 CG1 
                 VAL 
                 E 
                 723 
                 −16.163 
                 −1.892 
                 136.926 
                 1.00 
                 28.90 
                 E 
                 C 
               
               
                 ATOM 
                 27168 
                 CG2 
                 VAL 
                 E 
                 723 
                 −13.856 
                 −1.141 
                 137.461 
                 1.00 
                 28.65 
                 E 
                 C 
               
               
                 ATOM 
                 27169 
                 C 
                 VAL 
                 E 
                 723 
                 −15.006 
                 1.146 
                 138.959 
                 1.00 
                 51.77 
                 E 
                 C 
               
               
                 ATOM 
                 27170 
                 O 
                 VAL 
                 E 
                 723 
                 −15.462 
                 1.340 
                 140.083 
                 1.00 
                 53.93 
                 E 
                 O 
               
               
                 ATOM 
                 27171 
                 N 
                 SER 
                 E 
                 724 
                 −13.801 
                 1.570 
                 138.593 
                 1.00 
                 42.80 
                 E 
                 N 
               
               
                 ATOM 
                 27172 
                 CA 
                 SER 
                 E 
                 724 
                 −12.931 
                 2.294 
                 139.514 
                 1.00 
                 42.60 
                 E 
                 C 
               
               
                 ATOM 
                 27173 
                 CB 
                 SER 
                 E 
                 724 
                 −11.799 
                 2.985 
                 138.750 
                 1.00 
                 57.07 
                 E 
                 C 
               
               
                 ATOM 
                 27174 
                 OG 
                 SER 
                 E 
                 724 
                 −10.980 
                 2.038 
                 138.086 
                 1.00 
                 61.04 
                 E 
                 O 
               
               
                 ATOM 
                 27175 
                 C 
                 SER 
                 E 
                 724 
                 −13.716 
                 3.319 
                 140.328 
                 1.00 
                 40.83 
                 E 
                 C 
               
               
                 ATOM 
                 27176 
                 O 
                 SER 
                 E 
                 724 
                 −13.568 
                 3.402 
                 141.547 
                 1.00 
                 40.54 
                 E 
                 O 
               
               
                 ATOM 
                 27185 
                 N 
                 ASN 
                 E 
                 726 
                 −13.057 
                 6.538 
                 139.474 
                 1.00 
                 40.23 
                 E 
                 N 
               
               
                 ATOM 
                 27186 
                 CA 
                 ASN 
                 E 
                 726 
                 −11.943 
                 7.459 
                 139.631 
                 1.00 
                 39.80 
                 E 
                 C 
               
               
                 ATOM 
                 27187 
                 CB 
                 ASN 
                 E 
                 726 
                 −10.617 
                 6.730 
                 139.390 
                 1.00 
                 40.26 
                 E 
                 C 
               
               
                 ATOM 
                 27188 
                 CG 
                 ASN 
                 E 
                 726 
                 −10.408 
                 6.343 
                 137.928 
                 1.00 
                 42.08 
                 E 
                 C 
               
               
                 ATOM 
                 27189 
                 OD1 
                 ASN 
                 E 
                 726 
                 −11.345 
                 6.345 
                 137.128 
                 1.00 
                 42.97 
                 E 
                 O 
               
               
                 ATOM 
                 27190 
                 ND2 
                 ASN 
                 E 
                 726 
                 −9.171 
                 5.988 
                 137.581 
                 1.00 
                 42.90 
                 E 
                 N 
               
               
                 ATOM 
                 27191 
                 C 
                 ASN 
                 E 
                 726 
                 −12.031 
                 8.644 
                 138.698 
                 1.00 
                 40.02 
                 E 
                 C 
               
               
                 ATOM 
                 27192 
                 O 
                 ASN 
                 E 
                 726 
                 −11.018 
                 9.075 
                 138.144 
                 1.00 
                 41.23 
                 E 
                 O 
               
               
                 ATOM 
                 27499 
                 N 
                 GLN 
                 E 
                 767 
                 −10.146 
                 −4.183 
                 142.154 
                 1.00 
                 56.79 
                 E 
                 N 
               
               
                 ATOM 
                 27500 
                 CA 
                 GLN 
                 E 
                 767 
                 −9.698 
                 −3.192 
                 141.181 
                 1.00 
                 54.80 
                 E 
                 C 
               
               
                 ATOM 
                 27501 
                 CB 
                 GLN 
                 E 
                 767 
                 −10.398 
                 −3.412 
                 139.845 
                 1.00 
                 48.75 
                 E 
                 C 
               
               
                 ATOM 
                 27502 
                 CG 
                 GLN 
                 E 
                 767 
                 −9.861 
                 −4.597 
                 139.078 
                 1.00 
                 48.20 
                 E 
                 C 
               
               
                 ATOM 
                 27503 
                 CD 
                 GLN 
                 E 
                 767 
                 −10.422 
                 −4.699 
                 137.676 
                 1.00 
                 47.57 
                 E 
                 C 
               
               
                 ATOM 
                 27504 
                 OE1 
                 GLN 
                 E 
                 767 
                 −9.963 
                 −5.513 
                 136.875 
                 1.00 
                 46.04 
                 E 
                 O 
               
               
                 ATOM 
                 27505 
                 NE2 
                 GLN 
                 E 
                 767 
                 −11.420 
                 −3.874 
                 137.369 
                 1.00 
                 47.00 
                 E 
                 N 
               
               
                 ATOM 
                 27506 
                 C 
                 GLN 
                 E 
                 767 
                 −9.943 
                 −1.766 
                 141.637 
                 1.00 
                 54.64 
                 E 
                 C 
               
               
                 ATOM 
                 27507 
                 O 
                 GLN 
                 E 
                 767 
                 −10.456 
                 −1.539 
                 142.731 
                 1.00 
                 55.62 
                 E 
                 O 
               
               
                 ATOM 
                 27508 
                 N 
                 GLY 
                 E 
                 768 
                 −9.567 
                 −0.807 
                 140.789 
                 1.00 
                 50.62 
                 E 
                 N 
               
               
                 ATOM 
                 27509 
                 CA 
                 GLY 
                 E 
                 768 
                 −9.761 
                 0.603 
                 141.104 
                 1.00 
                 49.39 
                 E 
                 C 
               
               
                 ATOM 
                 27510 
                 C 
                 GLY 
                 E 
                 768 
                 −8.555 
                 1.518 
                 140.898 
                 1.00 
                 48.56 
                 E 
                 C 
               
               
                 ATOM 
                 27511 
                 O 
                 GLY 
                 E 
                 768 
                 −8.613 
                 2.706 
                 141.220 
                 1.00 
                 47.27 
                 E 
                 O 
               
               
                 ATOM 
                 27512 
                 N 
                 GLY 
                 E 
                 769 
                 −7.467 
                 0.981 
                 140.355 
                 1.00 
                 48.44 
                 E 
                 N 
               
               
                 ATOM 
                 27513 
                 CA 
                 GLY 
                 E 
                 769 
                 −6.288 
                 1.796 
                 140.157 
                 1.00 
                 48.52 
                 E 
                 C 
               
               
                 ATOM 
                 27514 
                 C 
                 GLY 
                 E 
                 769 
                 −5.882 
                 2.430 
                 141.478 
                 1.00 
                 48.98 
                 E 
                 C 
               
               
                 ATOM 
                 27515 
                 O 
                 GLY 
                 E 
                 769 
                 −5.939 
                 1.784 
                 142.517 
                 1.00 
                 50.43 
                 E 
                 O 
               
               
                 ATOM 
                 27966 
                 N 
                 GLU 
                 E 
                 827 
                 −5.497 
                 −2.722 
                 144.176 
                 1.00 
                 54.56 
                 E 
                 N 
               
               
                 ATOM 
                 27967 
                 CA 
                 GLU 
                 E 
                 827 
                 −4.381 
                 −2.981 
                 143.246 
                 1.00 
                 54.30 
                 E 
                 C 
               
               
                 ATOM 
                 27968 
                 CB 
                 GLU 
                 E 
                 827 
                 −4.786 
                 −2.716 
                 141.791 
                 1.00 
                 58.97 
                 E 
                 C 
               
               
                 ATOM 
                 27969 
                 CG 
                 GLU 
                 E 
                 827 
                 −5.933 
                 −3.497 
                 141.202 
                 1.00 
                 58.82 
                 E 
                 C 
               
               
                 ATOM 
                 27970 
                 CD 
                 GLU 
                 E 
                 827 
                 −6.282 
                 −2.979 
                 139.813 
                 1.00 
                 59.12 
                 E 
                 C 
               
               
                 ATOM 
                 27971 
                 OE1 
                 GLU 
                 E 
                 827 
                 −6.551 
                 −1.765 
                 139.679 
                 1.00 
                 59.54 
                 E 
                 O 
               
               
                 ATOM 
                 27972 
                 OE2 
                 GLU 
                 E 
                 827 
                 −6.284 
                 −3.776 
                 138.856 
                 1.00 
                 59.50 
                 E 
                 O 
               
               
                 ATOM 
                 27973 
                 C 
                 GLU 
                 E 
                 827 
                 −3.151 
                 −2.110 
                 143.478 
                 1.00 
                 54.38 
                 E 
                 C 
               
               
                 ATOM 
                 27974 
                 O 
                 GLU 
                 E 
                 827 
                 −2.012 
                 −2.573 
                 143.396 
                 1.00 
                 54.67 
                 E 
                 O 
               
               
                 ATOM 
                 28199 
                 N 
                 HIS 
                 E 
                 859 
                 −10.672 
                 −9.309 
                 135.849 
                 1.00 
                 43.96 
                 E 
                 N 
               
               
                 ATOM 
                 28200 
                 CA 
                 HIS 
                 E 
                 859 
                 −11.599 
                 −8.642 
                 134.955 
                 1.00 
                 42.22 
                 E 
                 C 
               
               
                 ATOM 
                 28201 
                 CB 
                 HIS 
                 E 
                 859 
                 −11.156 
                 −8.851 
                 133.499 
                 1.00 
                 44.60 
                 E 
                 C 
               
               
                 ATOM 
                 28202 
                 CG 
                 HIS 
                 E 
                 859 
                 −10.057 
                 −7.924 
                 133.079 
                 1.00 
                 44.48 
                 E 
                 C 
               
               
                 ATOM 
                 28203 
                 CD2 
                 HIS 
                 E 
                 859 
                 −10.091 
                 −6.757 
                 132.392 
                 1.00 
                 44.35 
                 E 
                 C 
               
               
                 ATOM 
                 28204 
                 ND1 
                 HIS 
                 E 
                 859 
                 −8.745 
                 −8.101 
                 133.465 
                 1.00 
                 44.66 
                 E 
                 N 
               
               
                 ATOM 
                 28205 
                 CE1 
                 HIS 
                 E 
                 859 
                 −8.018 
                 −7.083 
                 133.036 
                 1.00 
                 43.57 
                 E 
                 C 
               
               
                 ATOM 
                 28206 
                 NE2 
                 HIS 
                 E 
                 859 
                 −8.811 
                 −6.254 
                 132.382 
                 1.00 
                 44.22 
                 E 
                 N 
               
               
                 ATOM 
                 28207 
                 C 
                 HIS 
                 E 
                 859 
                 −13.085 
                 −8.919 
                 135.109 
                 1.00 
                 40.45 
                 E 
                 C 
               
               
                 ATOM 
                 28208 
                 O 
                 HIS 
                 E 
                 859 
                 −13.900 
                 −8.225 
                 134.499 
                 1.00 
                 39.42 
                 E 
                 O 
               
               
                 ATOM 
                 28225 
                 N 
                 GLN 
                 E 
                 862 
                 −15.810 
                 −6.234 
                 136.010 
                 1.00 
                 45.03 
                 E 
                 N 
               
               
                 ATOM 
                 28226 
                 CA 
                 GLN 
                 E 
                 862 
                 −16.440 
                 −5.329 
                 135.047 
                 1.00 
                 44.85 
                 E 
                 C 
               
               
                 ATOM 
                 28227 
                 CB 
                 GLN 
                 E 
                 862 
                 −15.731 
                 −5.376 
                 133.706 
                 1.00 
                 41.80 
                 E 
                 C 
               
               
                 ATOM 
                 28228 
                 CG 
                 GLN 
                 E 
                 862 
                 −14.429 
                 −4.627 
                 133.689 
                 1.00 
                 42.36 
                 E 
                 C 
               
               
                 ATOM 
                 28229 
                 CD 
                 GLN 
                 E 
                 862 
                 −13.614 
                 −4.941 
                 132.457 
                 1.00 
                 43.95 
                 E 
                 C 
               
               
                 ATOM 
                 28230 
                 OE1 
                 GLN 
                 E 
                 862 
                 −12.451 
                 −4.541 
                 132.350 
                 1.00 
                 45.62 
                 E 
                 O 
               
               
                 ATOM 
                 28231 
                 NE2 
                 GLN 
                 E 
                 862 
                 −14.218 
                 −5.672 
                 131.511 
                 1.00 
                 43.67 
                 E 
                 N 
               
               
                 ATOM 
                 28232 
                 C 
                 GLN 
                 E 
                 862 
                 −17.899 
                 −5.688 
                 134.849 
                 1.00 
                 46.08 
                 E 
                 C 
               
               
                 ATOM 
                 28233 
                 O 
                 GLN 
                 E 
                 862 
                 −18.715 
                 −4.830 
                 134.514 
                 1.00 
                 45.77 
                 E 
                 O 
               
               
                 ATOM 
                 28726 
                 N 
                 ARG 
                 E 
                 952 
                 −4.527 
                 9.341 
                 138.541 
                 1.00 
                 46.53 
                 E 
                 N 
               
               
                 ATOM 
                 28727 
                 CA 
                 ARG 
                 E 
                 952 
                 −5.670 
                 9.279 
                 137.643 
                 1.00 
                 42.59 
                 E 
                 C 
               
               
                 ATOM 
                 28728 
                 CB 
                 ARG 
                 E 
                 952 
                 −5.553 
                 8.062 
                 136.727 
                 1.00 
                 52.48 
                 E 
                 C 
               
               
                 ATOM 
                 28729 
                 CG 
                 ARG 
                 E 
                 952 
                 −4.333 
                 8.078 
                 135.821 
                 1.00 
                 51.19 
                 E 
                 C 
               
               
                 ATOM 
                 28730 
                 CD 
                 ARG 
                 E 
                 952 
                 −4.202 
                 6.786 
                 135.033 
                 1.00 
                 50.26 
                 E 
                 C 
               
               
                 ATOM 
                 28731 
                 NE 
                 ARG 
                 E 
                 952 
                 −5.407 
                 6.464 
                 134.270 
                 1.00 
                 50.19 
                 E 
                 N 
               
               
                 ATOM 
                 28732 
                 CZ 
                 ARG 
                 E 
                 952 
                 −6.420 
                 5.732 
                 134.722 
                 1.00 
                 49.56 
                 E 
                 C 
               
               
                 ATOM 
                 28733 
                 NH1 
                 ARG 
                 E 
                 952 
                 −6.393 
                 5.231 
                 135.944 
                 1.00 
                 50.34 
                 E 
                 N 
               
               
                 ATOM 
                 28734 
                 NH2 
                 ARG 
                 E 
                 952 
                 −7.465 
                 5.500 
                 133.949 
                 1.00 
                 49.07 
                 E 
                 N 
               
               
                 ATOM 
                 28735 
                 C 
                 ARG 
                 E 
                 952 
                 −6.956 
                 9.179 
                 138.431 
                 1.00 
                 40.06 
                 E 
                 C 
               
               
                 ATOM 
                 28736 
                 O 
                 ARG 
                 E 
                 952 
                 −7.740 
                 8.267 
                 138.218 
                 1.00 
                 40.57 
                 E 
                 O 
               
               
                 ATOM 
                 46526 
                 O2P 
                 FDP 
                 P 
                 5 
                 −8.320 
                 3.029 
                 136.143 
                 1.00 
                 64.30 
                 P 
                 O 
               
               
                 ATOM 
                 46527 
                 P1 
                 FDP 
                 P 
                 5 
                 −9.484 
                 2.294 
                 135.308 
                 1.00 
                 64.97 
                 P 
                 P 
               
               
                 ATOM 
                 46528 
                 O3P 
                 FDP 
                 P 
                 5 
                 −9.695 
                 3.178 
                 133.979 
                 1.00 
                 64.42 
                 P 
                 O 
               
               
                 ATOM 
                 46529 
                 O1P 
                 FDP 
                 P 
                 5 
                 −10.741 
                 2.182 
                 136.083 
                 1.00 
                 65.03 
                 P 
                 O 
               
               
                 ATOM 
                 46530 
                 O2 
                 FDP 
                 P 
                 5 
                 −8.887 
                 0.876 
                 134.827 
                 1.00 
                 64.82 
                 P 
                 O 
               
               
                 ATOM 
                 46531 
                 C2 
                 FDP 
                 P 
                 5 
                 −8.854 
                 −0.294 
                 135.658 
                 1.00 
                 64.30 
                 P 
                 C 
               
               
                 ATOM 
                 46532 
                 C1 
                 FDP 
                 P 
                 5 
                 −8.491 
                 0.074 
                 137.100 
                 1.00 
                 63.69 
                 P 
                 C 
               
               
                 ATOM 
                 46533 
                 O1 
                 FDP 
                 P 
                 5 
                 −8.454 
                 −1.102 
                 137.912 
                 1.00 
                 60.73 
                 P 
                 O 
               
               
                 ATOM 
                 46534 
                 O5 
                 FDP 
                 P 
                 5 
                 −7.846 
                 −1.171 
                 135.132 
                 1.00 
                 64.50 
                 P 
                 O 
               
               
                 ATOM 
                 46535 
                 C3 
                 FDP 
                 P 
                 5 
                 −10.174 
                 −1.073 
                 135.604 
                 1.00 
                 65.04 
                 P 
                 C 
               
               
                 ATOM 
                 46536 
                 O3 
                 FDP 
                 P 
                 5 
                 −11.251 
                 −0.224 
                 135.196 
                 1.00 
                 64.96 
                 P 
                 O 
               
               
                 ATOM 
                 46537 
                 C4 
                 FDP 
                 P 
                 5 
                 −9.882 
                 −2.089 
                 134.503 
                 1.00 
                 64.39 
                 P 
                 C 
               
               
                 ATOM 
                 46538 
                 O4 
                 FDP 
                 P 
                 5 
                 −10.747 
                 −3.220 
                 134.650 
                 1.00 
                 63.58 
                 P 
                 O 
               
               
                 ATOM 
                 46539 
                 C5 
                 FDP 
                 P 
                 5 
                 −8.454 
                 −2.454 
                 134.903 
                 1.00 
                 64.67 
                 P 
                 C 
               
               
                 ATOM 
                 46540 
                 C6 
                 FDP 
                 P 
                 5 
                 −7.727 
                 −3.266 
                 133.832 
                 1.00 
                 65.81 
                 P 
                 C 
               
               
                 ATOM 
                 46541 
                 O6 
                 FDP 
                 P 
                 5 
                 −7.767 
                 −2.603 
                 132.564 
                 1.00 
                 67.34 
                 P 
                 O 
               
               
                 ATOM 
                 46542 
                 P2 
                 FDP 
                 P 
                 5 
                 −7.026 
                 −3.240 
                 131.285 
                 1.00 
                 68.65 
                 P 
                 P 
               
               
                 ATOM 
                 46543 
                 O5P 
                 FDP 
                 P 
                 5 
                 −6.305 
                 −4.587 
                 131.796 
                 1.00 
                 67.22 
                 P 
                 O 
               
               
                 ATOM 
                 46544 
                 O6P 
                 FDP 
                 P 
                 5 
                 −8.223 
                 −3.684 
                 130.301 
                 1.00 
                 68.83 
                 P 
                 O 
               
               
                 ATOM 
                 46545 
                 O4P 
                 FDP 
                 P 
                 5 
                 −6.078 
                 −2.304 
                 130.641 
                 1.00 
                 67.08 
                 P 
                 O 
               
               
                 END 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 1b 
               
               
                   
               
             
            
               
                 ATOM 
                 4238 
                 N 
                 ARG 
                 A 
                 760 
                 19.077 
                 71.248 
                 89.275 
                 1.00 
                 66.25 
                 A 
                 N 
               
               
                 ATOM 
                 4239 
                 CA 
                 ARG 
                 A 
                 760 
                 18.203 
                 70.781 
                 90.326 
                 1.00 
                 64.93 
                 A 
                 C 
               
               
                 ATOM 
                 4240 
                 CB 
                 ARG 
                 A 
                 760 
                 16.781 
                 71.291 
                 90.071 
                 1.00 
                 51.29 
                 A 
                 C 
               
               
                 ATOM 
                 4241 
                 CG 
                 ARG 
                 A 
                 760 
                 15.726 
                 70.610 
                 90.902 
                 1.00 
                 50.50 
                 A 
                 C 
               
               
                 ATOM 
                 4242 
                 CD 
                 ARG 
                 A 
                 760 
                 14.391 
                 71.303 
                 90.765 
                 1.00 
                 49.71 
                 A 
                 C 
               
               
                 ATOM 
                 4243 
                 NE 
                 ARG 
                 A 
                 760 
                 14.395 
                 72.656 
                 91.328 
                 1.00 
                 47.57 
                 A 
                 N 
               
               
                 ATOM 
                 4244 
                 CZ 
                 ARG 
                 A 
                 760 
                 13.284 
                 73.355 
                 91.539 
                 1.00 
                 47.02 
                 A 
                 C 
               
               
                 ATOM 
                 4245 
                 NH1 
                 ARG 
                 A 
                 760 
                 12.113 
                 72.816 
                 91.237 
                 1.00 
                 47.43 
                 A 
                 N 
               
               
                 ATOM 
                 4246 
                 NH2 
                 ARG 
                 A 
                 760 
                 13.330 
                 74.584 
                 92.030 
                 1.00 
                 46.43 
                 A 
                 N 
               
               
                 ATOM 
                 4247 
                 C 
                 ARG 
                 A 
                 760 
                 18.233 
                 69.261 
                 90.420 
                 1.00 
                 64.58 
                 A 
                 C 
               
               
                 ATOM 
                 4248 
                 O 
                 ARG 
                 A 
                 760 
                 18.213 
                 68.554 
                 89.405 
                 1.00 
                 64.15 
                 A 
                 O 
               
               
                 ATOM 
                 4954 
                 N 
                 ARG 
                 A 
                 853 
                 14.782 
                 68.630 
                 95.568 
                 1.00 
                 44.54 
                 A 
                 N 
               
               
                 ATOM 
                 4955 
                 CA 
                 ARG 
                 A 
                 853 
                 13.363 
                 68.336 
                 95.685 
                 1.00 
                 46.94 
                 A 
                 C 
               
               
                 ATOM 
                 4956 
                 CB 
                 ARG 
                 A 
                 853 
                 12.608 
                 68.673 
                 94.388 
                 1.00 
                 38.58 
                 A 
                 C 
               
               
                 ATOM 
                 4957 
                 CG 
                 ARG 
                 A 
                 853 
                 12.391 
                 70.171 
                 94.140 
                 1.00 
                 35.99 
                 A 
                 C 
               
               
                 ATOM 
                 4958 
                 CD 
                 ARG 
                 A 
                 853 
                 11.439 
                 70.806 
                 95.154 
                 1.00 
                 33.30 
                 A 
                 C 
               
               
                 ATOM 
                 4959 
                 NE 
                 ARG 
                 A 
                 853 
                 11.210 
                 72.236 
                 94.918 
                 1.00 
                 30.21 
                 A 
                 N 
               
               
                 ATOM 
                 4960 
                 CZ 
                 ARG 
                 A 
                 853 
                 12.129 
                 73.179 
                 95.091 
                 1.00 
                 27.44 
                 A 
                 C 
               
               
                 ATOM 
                 4961 
                 NH1 
                 ARG 
                 A 
                 853 
                 13.337 
                 72.838 
                 95.497 
                 1.00 
                 28.12 
                 A 
                 N 
               
               
                 ATOM 
                 4962 
                 NH2 
                 ARG 
                 A 
                 853 
                 11.844 
                 74.460 
                 94.879 
                 1.00 
                 24.54 
                 A 
                 N 
               
               
                 ATOM 
                 4963 
                 C 
                 ARG 
                 A 
                 853 
                 13.197 
                 66.868 
                 96.025 
                 1.00 
                 49.22 
                 A 
                 C 
               
               
                 ATOM 
                 4964 
                 O 
                 ARG 
                 A 
                 853 
                 14.086 
                 66.045 
                 95.791 
                 1.00 
                 48.57 
                 A 
                 O 
               
               
                 ATOM 
                 8816 
                 N 
                 ALA 
                 B 
                 596 
                 7.520 
                 78.037 
                 86.751 
                 1.00 
                 42.66 
                 B 
                 N 
               
               
                 ATOM 
                 8817 
                 CA 
                 ALA 
                 B 
                 596 
                 7.790 
                 76.616 
                 86.760 
                 1.00 
                 41.63 
                 B 
                 C 
               
               
                 ATOM 
                 8818 
                 C 
                 ALA 
                 B 
                 596 
                 6.962 
                 75.944 
                 87.839 
                 1.00 
                 52.25 
                 B 
                 C 
               
               
                 ATOM 
                 8819 
                 C 
                 ALA 
                 B 
                 596 
                 7.463 
                 76.020 
                 85.407 
                 1.00 
                 41.31 
                 B 
                 C 
               
               
                 ATOM 
                 8820 
                 O 
                 ALA 
                 B 
                 596 
                 6.757 
                 76.623 
                 84.602 
                 1.00 
                 41.28 
                 B 
                 O 
               
               
                 ATOM 
                 9289 
                 N 
                 ARG 
                 B 
                 658 
                 12.243 
                 81.331 
                 88.859 
                 1.00 
                 55.29 
                 B 
                 N 
               
               
                 ATOM 
                 9290 
                 CA 
                 ARG 
                 B 
                 658 
                 11.252 
                 82.230 
                 89.436 
                 1.00 
                 54.74 
                 B 
                 C 
               
               
                 ATOM 
                 9291 
                 CB 
                 ARG 
                 B 
                 658 
                 10.670 
                 81.612 
                 90.727 
                 1.00 
                 45.12 
                 B 
                 C 
               
               
                 ATOM 
                 9292 
                 CG 
                 ARG 
                 B 
                 658 
                 9.703 
                 80.429 
                 90.531 
                 1.00 
                 43.86 
                 B 
                 C 
               
               
                 ATOM 
                 9293 
                 CD 
                 ARG 
                 B 
                 658 
                 9.410 
                 79.734 
                 91.866 
                 1.00 
                 44.11 
                 B 
                 C 
               
               
                 ATOM 
                 9294 
                 NE 
                 ARG 
                 B 
                 658 
                 8.699 
                 78.459 
                 91.718 
                 1.00 
                 43.60 
                 B 
                 N 
               
               
                 ATOM 
                 9295 
                 CZ 
                 ARG 
                 B 
                 658 
                 7.391 
                 78.344 
                 91.489 
                 1.00 
                 43.55 
                 B 
                 C 
               
               
                 ATOM 
                 9296 
                 NH1 
                 ARG 
                 B 
                 658 
                 6.636 
                 79.433 
                 91.388 
                 1.00 
                 43.76 
                 B 
                 N 
               
               
                 ATOM 
                 9297 
                 NH2 
                 ARG 
                 B 
                 658 
                 6.841 
                 77.142 
                 91.348 
                 1.00 
                 41.28 
                 B 
                 N 
               
               
                 ATOM 
                 9298 
                 C 
                 ARG 
                 B 
                 658 
                 11.754 
                 83.635 
                 89.732 
                 1.00 
                 54.72 
                 B 
                 C 
               
               
                 ATOM 
                 9299 
                 O 
                 ARG 
                 B 
                 658 
                 10.983 
                 84.470 
                 90.208 
                 1.00 
                 55.44 
                 B 
                 O 
               
               
                 ATOM 
                 9532 
                 N 
                 GLU 
                 B 
                 688 
                 4.811 
                 84.215 
                 88.481 
                 1.00 
                 64.06 
                 B 
                 N 
               
               
                 ATOM 
                 9533 
                 CA 
                 GLU 
                 B 
                 688 
                 6.066 
                 84.927 
                 88.662 
                 1.00 
                 65.43 
                 B 
                 C 
               
               
                 ATOM 
                 9534 
                 CB 
                 GLU 
                 B 
                 688 
                 7.217 
                 83.927 
                 88.810 
                 1.00 
                 59.75 
                 B 
                 C 
               
               
                 ATOM 
                 9535 
                 CG 
                 GLU 
                 B 
                 688 
                 7.019 
                 82.962 
                 89.974 
                 1.00 
                 59.29 
                 B 
                 C 
               
               
                 ATOM 
                 9536 
                 CD 
                 GLU 
                 B 
                 688 
                 6.463 
                 83.662 
                 91.201 
                 1.00 
                 58.97 
                 B 
                 C 
               
               
                 ATOM 
                 9537 
                 OE1 
                 GLU 
                 B 
                 688 
                 6.875 
                 84.815 
                 91.472 
                 1.00 
                 58.33 
                 B 
                 O 
               
               
                 ATOM 
                 9538 
                 OE2 
                 GLU 
                 B 
                 688 
                 5.618 
                 83.060 
                 91.892 
                 1.00 
                 58.35 
                 B 
                 O 
               
               
                 ATOM 
                 9539 
                 C 
                 GLU 
                 B 
                 688 
                 6.251 
                 85.844 
                 87.451 
                 1.00 
                 65.83 
                 B 
                 C 
               
               
                 ATOM 
                 9540 
                 O 
                 GLU 
                 B 
                 688 
                 6.687 
                 86.985 
                 87.577 
                 1.00 
                 66.10 
                 B 
                 O 
               
               
                 ATOM 
                 9762 
                 N 
                 THR 
                 B 
                 716 
                 0.895 
                 77.642 
                 83.597 
                 1.00 
                 58.37 
                 B 
                 N 
               
               
                 ATOM 
                 9763 
                 CA 
                 THR 
                 B 
                 716 
                 1.199 
                 77.017 
                 84.881 
                 1.00 
                 56.38 
                 B 
                 C 
               
               
                 ATOM 
                 9764 
                 CB 
                 THR 
                 B 
                 716 
                 1.240 
                 78.061 
                 86.012 
                 1.00 
                 61.17 
                 B 
                 C 
               
               
                 ATOM 
                 9765 
                 OG1 
                 THR 
                 B 
                 716 
                 0.883 
                 77.443 
                 87.250 
                 1.00 
                 61.30 
                 B 
                 O 
               
               
                 ATOM 
                 9766 
                 CG2 
                 THR 
                 B 
                 716 
                 0.252 
                 79.171 
                 85.749 
                 1.00 
                 62.23 
                 B 
                 C 
               
               
                 ATOM 
                 9767 
                 C 
                 THR 
                 B 
                 716 
                 0.063 
                 76.037 
                 85.150 
                 1.00 
                 55.58 
                 B 
                 C 
               
               
                 ATOM 
                 9768 
                 O 
                 THR 
                 B 
                 716 
                 −1.091 
                 76.333 
                 84.846 
                 1.00 
                 55.90 
                 B 
                 O 
               
               
                 ATOM 
                 9769 
                 N 
                 LEU 
                 B 
                 717 
                 0.377 
                 74.873 
                 85.707 
                 1.00 
                 58.49 
                 B 
                 N 
               
               
                 ATOM 
                 9770 
                 CA 
                 LEU 
                 B 
                 717 
                 −0.650 
                 73.871 
                 85.990 
                 1.00 
                 56.82 
                 B 
                 C 
               
               
                 ATOM 
                 9771 
                 CB 
                 LEU 
                 B 
                 717 
                 0.030 
                 72.565 
                 86.429 
                 1.00 
                 46.67 
                 B 
                 C 
               
               
                 ATOM 
                 9772 
                 CG 
                 LEU 
                 B 
                 717 
                 1.080 
                 72.637 
                 87.546 
                 1.00 
                 47.47 
                 B 
                 C 
               
               
                 ATOM 
                 9773 
                 CD1 
                 LEU 
                 B 
                 717 
                 0.367 
                 72.956 
                 88.843 
                 1.00 
                 47.98 
                 B 
                 C 
               
               
                 ATOM 
                 9774 
                 CD2 
                 LEU 
                 B 
                 717 
                 1.841 
                 71.306 
                 87.687 
                 1.00 
                 47.27 
                 B 
                 C 
               
               
                 ATOM 
                 9775 
                 C 
                 LEU 
                 B 
                 717 
                 −1.704 
                 74.341 
                 87.026 
                 1.00 
                 55.29 
                 B 
                 C 
               
               
                 ATOM 
                 9776 
                 O 
                 LEU 
                 B 
                 717 
                 −2.861 
                 73.912 
                 87.000 
                 1.00 
                 54.28 
                 B 
                 O 
               
               
                 ATOM 
                 9777 
                 N 
                 SER 
                 B 
                 718 
                 −1.296 
                 75.244 
                 87.911 
                 1.00 
                 41.68 
                 B 
                 N 
               
               
                 ATOM 
                 9778 
                 CA 
                 SER 
                 B 
                 718 
                 −2.168 
                 75.781 
                 88.945 
                 1.00 
                 42.21 
                 B 
                 C 
               
               
                 ATOM 
                 9779 
                 CB 
                 SER 
                 B 
                 718 
                 −1.366 
                 76.674 
                 89.892 
                 1.00 
                 57.27 
                 B 
                 C 
               
               
                 ATOM 
                 9780 
                 OG 
                 SER 
                 B 
                 718 
                 −0.140 
                 76.082 
                 90.265 
                 1.00 
                 57.44 
                 B 
                 O 
               
               
                 ATOM 
                 9781 
                 C 
                 SER 
                 B 
                 718 
                 −3.333 
                 76.605 
                 88.391 
                 1.00 
                 43.08 
                 B 
                 C 
               
               
                 ATOM 
                 9782 
                 O 
                 SER 
                 B 
                 718 
                 −4.441 
                 76.544 
                 88.922 
                 1.00 
                 43.24 
                 B 
                 O 
               
               
                 ATOM 
                 9791 
                 N 
                 ASN 
                 B 
                 720 
                 −3.278 
                 79.928 
                 88.251 
                 1.00 
                 63.03 
                 B 
                 N 
               
               
                 ATOM 
                 9792 
                 CA 
                 ASN 
                 B 
                 720 
                 −3.378 
                 81.000 
                 89.231 
                 1.00 
                 64.27 
                 B 
                 C 
               
               
                 ATOM 
                 9793 
                 CB 
                 ASN 
                 B 
                 720 
                 −2.435 
                 80.722 
                 90.424 
                 1.00 
                 56.96 
                 B 
                 C 
               
               
                 ATOM 
                 9794 
                 CG 
                 ASN 
                 B 
                 720 
                 −0.949 
                 80.713 
                 90.042 
                 1.00 
                 56.53 
                 B 
                 C 
               
               
                 ATOM 
                 9795 
                 OD1 
                 ASN 
                 B 
                 720 
                 −0.527 
                 80.050 
                 89.087 
                 1.00 
                 56.28 
                 B 
                 O 
               
               
                 ATOM 
                 9796 
                 ND2 
                 ASN 
                 B 
                 720 
                 −0.149 
                 81.434 
                 90.813 
                 1.00 
                 56.39 
                 B 
                 N 
               
               
                 ATOM 
                 9797 
                 C 
                 ASN 
                 B 
                 720 
                 −3.117 
                 82.394 
                 88.678 
                 1.00 
                 65.24 
                 B 
                 C 
               
               
                 ATOM 
                 9798 
                 O 
                 ASN 
                 B 
                 720 
                 −3.250 
                 83.382 
                 89.401 
                 1.00 
                 65.17 
                 B 
                 O 
               
               
                 ATOM 
                 10093 
                 N 
                 GLN 
                 B 
                 761 
                 −1.224 
                 69.548 
                 93.756 
                 1.00 
                 49.38 
                 B 
                 N 
               
               
                 ATOM 
                 10094 
                 CA 
                 GLN 
                 B 
                 761 
                 −0.541 
                 70.829 
                 93.565 
                 1.00 
                 49.08 
                 B 
                 C 
               
               
                 ATOM 
                 10095 
                 CB 
                 GLN 
                 B 
                 761 
                 0.507 
                 70.726 
                 92.458 
                 1.00 
                 48.74 
                 B 
                 C 
               
               
                 ATOM 
                 10096 
                 CG 
                 GLN 
                 B 
                 761 
                 1.857 
                 70.217 
                 92.924 
                 1.00 
                 50.25 
                 B 
                 C 
               
               
                 ATOM 
                 10097 
                 CD 
                 GLN 
                 B 
                 761 
                 2.861 
                 70.060 
                 91.789 
                 1.00 
                 51.04 
                 B 
                 C 
               
               
                 ATOM 
                 10098 
                 OE1 
                 GLN 
                 B 
                 761 
                 3.986 
                 69.612 
                 92.003 
                 1.00 
                 51.31 
                 B 
                 O 
               
               
                 ATOM 
                 10099 
                 NE2 
                 GLN 
                 B 
                 761 
                 2.455 
                 70.422 
                 90.577 
                 1.00 
                 52.66 
                 B 
                 N 
               
               
                 ATOM 
                 10100 
                 C 
                 GLN 
                 B 
                 761 
                 −1.504 
                 71.950 
                 93.218 
                 1.00 
                 48.43 
                 B 
                 C 
               
               
                 ATOM 
                 10101 
                 O 
                 GLN 
                 B 
                 761 
                 −2.705 
                 71.721 
                 93.060 
                 1.00 
                 49.19 
                 B 
                 O 
               
               
                 ATOM 
                 10102 
                 N 
                 GLY 
                 B 
                 762 
                 −0.973 
                 73.162 
                 93.101 
                 1.00 
                 38.22 
                 B 
                 N 
               
               
                 ATOM 
                 10103 
                 CA 
                 GLY 
                 B 
                 762 
                 −1.806 
                 74.292 
                 92.764 
                 1.00 
                 37.62 
                 B 
                 C 
               
               
                 ATOM 
                 10104 
                 C 
                 GLY 
                 B 
                 762 
                 −1.600 
                 75.500 
                 93.655 
                 1.00 
                 38.70 
                 B 
                 C 
               
               
                 ATOM 
                 10105 
                 O 
                 GLY 
                 B 
                 762 
                 −2.200 
                 76.553 
                 93.445 
                 1.00 
                 40.36 
                 B 
                 O 
               
               
                 ATOM 
                 10106 
                 N 
                 GLY 
                 B 
                 763 
                 −0.745 
                 75.383 
                 94.653 
                 1.00 
                 39.55 
                 B 
                 N 
               
               
                 ATOM 
                 10107 
                 CA 
                 GLY 
                 B 
                 763 
                 −0.547 
                 76.526 
                 95.516 
                 1.00 
                 40.11 
                 B 
                 C 
               
               
                 ATOM 
                 10108 
                 C 
                 GLY 
                 B 
                 763 
                 −1.871 
                 76.897 
                 96.154 
                 1.00 
                 42.01 
                 B 
                 C 
               
               
                 ATOM 
                 10109 
                 O 
                 GLY 
                 B 
                 763 
                 −2.561 
                 76.034 
                 96.707 
                 1.00 
                 43.43 
                 B 
                 O 
               
               
                 ATOM 
                 10546 
                 N 
                 THR 
                 B 
                 821 
                 −2.601 
                 70.694 
                 98.759 
                 1.00 
                 42.22 
                 B 
                 N 
               
               
                 ATOM 
                 10547 
                 CA 
                 THR 
                 B 
                 821 
                 −1.712 
                 70.944 
                 99.891 
                 1.00 
                 39.50 
                 B 
                 C 
               
               
                 ATOM 
                 10548 
                 CB 
                 THR 
                 B 
                 821 
                 −0.258 
                 71.108 
                 99.425 
                 1.00 
                 20.00 
                 B 
                 C 
               
               
                 ATOM 
                 10549 
                 OG1 
                 THR 
                 B 
                 821 
                 0.101 
                 69.985 
                 98.616 
                 1.00 
                 20.98 
                 B 
                 O 
               
               
                 ATOM 
                 10550 
                 CG2 
                 THR 
                 B 
                 821 
                 0.690 
                 71.209 
                 100.612 
                 1.00 
                 15.44 
                 B 
                 C 
               
               
                 ATOM 
                 10551 
                 C 
                 THR 
                 B 
                 821 
                 −2.056 
                 72.175 
                 100.719 
                 1.00 
                 40.64 
                 B 
                 C 
               
               
                 ATOM 
                 10552 
                 O 
                 THR 
                 B 
                 821 
                 −2.080 
                 72.108 
                 101.944 
                 1.00 
                 41.93 
                 B 
                 O 
               
               
                 ATOM 
                 10775 
                 N 
                 HIS 
                 B 
                 853 
                 5.907 
                 66.188 
                 91.557 
                 1.00 
                 38.36 
                 B 
                 N 
               
               
                 ATOM 
                 10776 
                 CA 
                 HIS 
                 B 
                 853 
                 6.163 
                 66.916 
                 90.324 
                 1.00 
                 37.94 
                 B 
                 C 
               
               
                 ATOM 
                 10777 
                 CB 
                 HIS 
                 B 
                 853 
                 7.647 
                 67.308 
                 90.224 
                 1.00 
                 43.77 
                 B 
                 C 
               
               
                 ATOM 
                 10778 
                 CG 
                 HIS 
                 B 
                 853 
                 8.040 
                 68.441 
                 91.124 
                 1.00 
                 44.94 
                 B 
                 C 
               
               
                 ATOM 
                 10779 
                 CD2 
                 HIS 
                 B 
                 853 
                 8.405 
                 69.715 
                 90.845 
                 1.00 
                 45.52 
                 B 
                 C 
               
               
                 ATOM 
                 10780 
                 ND1 
                 HIS 
                 B 
                 853 
                 8.052 
                 68.336 
                 92.499 
                 1.00 
                 45.20 
                 B 
                 N 
               
               
                 ATOM 
                 10781 
                 CE1 
                 HIS 
                 B 
                 853 
                 8.405 
                 69.495 
                 93.026 
                 1.00 
                 45.33 
                 B 
                 C 
               
               
                 ATOM 
                 10782 
                 NE2 
                 HIS 
                 B 
                 853 
                 8.625 
                 70.349 
                 92.045 
                 1.00 
                 45.66 
                 B 
                 N 
               
               
                 ATOM 
                 10783 
                 C 
                 HIS 
                 B 
                 853 
                 5.734 
                 66.206 
                 89.051 
                 1.00 
                 38.02 
                 B 
                 C 
               
               
                 ATOM 
                 10784 
                 O 
                 HIS 
                 B 
                 853 
                 5.764 
                 66.807 
                 87.978 
                 1.00 
                 36.51 
                 B 
                 O 
               
               
                 ATOM 
                 10801 
                 N 
                 GLN 
                 B 
                 856 
                 3.392 
                 67.839 
                 86.576 
                 1.00 
                 45.51 
                 B 
                 N 
               
               
                 ATOM 
                 10802 
                 CA 
                 GLN 
                 B 
                 856 
                 3.628 
                 68.789 
                 85.476 
                 1.00 
                 46.12 
                 B 
                 C 
               
               
                 ATOM 
                 10803 
                 CB 
                 GLN 
                 B 
                 856 
                 4.991 
                 69.458 
                 85.617 
                 1.00 
                 36.58 
                 B 
                 C 
               
               
                 ATOM 
                 10804 
                 CG 
                 GLN 
                 B 
                 856 
                 5.161 
                 70.155 
                 86.948 
                 1.00 
                 36.05 
                 B 
                 C 
               
               
                 ATOM 
                 10805 
                 CD 
                 GLN 
                 B 
                 856 
                 6.503 
                 70.845 
                 87.096 
                 1.00 
                 35.54 
                 B 
                 C 
               
               
                 ATOM 
                 10806 
                 OE1 
                 GLN 
                 B 
                 856 
                 7.513 
                 70.400 
                 86.544 
                 1.00 
                 35.35 
                 B 
                 O 
               
               
                 ATOM 
                 10807 
                 NE2 
                 GLN 
                 B 
                 856 
                 6.525 
                 71.930 
                 87.862 
                 1.00 
                 34.45 
                 B 
                 N 
               
               
                 ATOM 
                 10808 
                 C 
                 GLN 
                 B 
                 856 
                 3.537 
                 68.014 
                 84.149 
                 1.00 
                 47.16 
                 B 
                 C 
               
               
                 ATOM 
                 10809 
                 O 
                 GLN 
                 B 
                 856 
                 3.406 
                 68.594 
                 83.065 
                 1.00 
                 47.21 
                 B 
                 O 
               
               
                 ATOM 
                 11401 
                 N 
                 ARG 
                 B 
                 935 
                 −0.953 
                 84.973 
                 95.475 
                 1.00 
                 54.95 
                 B 
                 N 
               
               
                 ATOM 
                 11402 
                 CA 
                 ARG 
                 B 
                 935 
                 −0.476 
                 84.735 
                 94.118 
                 1.00 
                 52.80 
                 B 
                 C 
               
               
                 ATOM 
                 11403 
                 CB 
                 ARG 
                 B 
                 935 
                 0.702 
                 83.765 
                 94.080 
                 1.00 
                 51.40 
                 B 
                 C 
               
               
                 ATOM 
                 11404 
                 CG 
                 ARG 
                 B 
                 935 
                 2.072 
                 84.404 
                 94.255 
                 1.00 
                 50.21 
                 B 
                 C 
               
               
                 ATOM 
                 11405 
                 CD 
                 ARG 
                 B 
                 935 
                 3.100 
                 83.562 
                 93.519 
                 1.00 
                 49.92 
                 B 
                 C 
               
               
                 ATOM 
                 11406 
                 NE 
                 ARG 
                 B 
                 935 
                 2.793 
                 82.156 
                 93.732 
                 1.00 
                 51.19 
                 B 
                 N 
               
               
                 ATOM 
                 11407 
                 CZ 
                 ARG 
                 B 
                 935 
                 3.207 
                 81.155 
                 92.965 
                 1.00 
                 51.30 
                 B 
                 C 
               
               
                 ATOM 
                 11408 
                 NH1 
                 ARG 
                 B 
                 935 
                 2.854 
                 79.907 
                 93.262 
                 1.00 
                 50.45 
                 B 
                 N 
               
               
                 ATOM 
                 11409 
                 NH2 
                 ARG 
                 B 
                 935 
                 3.968 
                 81.396 
                 91.912 
                 1.00 
                 51.06 
                 B 
                 N 
               
               
                 ATOM 
                 11410 
                 C 
                 ARG 
                 B 
                 935 
                 −1.590 
                 84.190 
                 93.249 
                 1.00 
                 52.62 
                 B 
                 C 
               
               
                 ATOM 
                 11411 
                 O 
                 ARG 
                 B 
                 935 
                 −1.413 
                 83.188 
                 92.553 
                 1.00 
                 52.78 
                 B 
                 O 
               
               
                 ATOM 
                 46466 
                 O2P 
                 FDP 
                 P 
                 2 
                 2.564 
                 77.669 
                 89.518 
                 1.00 
                 64.79 
                 P 
                 O 
               
               
                 ATOM 
                 46467 
                 P1 
                 FDP 
                 P 
                 2 
                 2.932 
                 77.562 
                 91.081 
                 1.00 
                 65.07 
                 P 
                 P 
               
               
                 ATOM 
                 46468 
                 O3P 
                 FDP 
                 P 
                 2 
                 1.908 
                 78.563 
                 91.816 
                 1.00 
                 64.74 
                 P 
                 O 
               
               
                 ATOM 
                 46469 
                 O1P 
                 FDP 
                 P 
                 2 
                 4.347 
                 77.907 
                 91.349 
                 1.00 
                 64.33 
                 P 
                 O 
               
               
                 ATOM 
                 46470 
                 O2 
                 FDP 
                 P 
                 2 
                 2.505 
                 76.087 
                 91.572 
                 1.00 
                 64.09 
                 P 
                 O 
               
               
                 ATOM 
                 46471 
                 C2 
                 FDP 
                 P 
                 2 
                 3.298 
                 74.903 
                 91.416 
                 1.00 
                 61.84 
                 P 
                 C 
               
               
                 ATOM 
                 46472 
                 C1 
                 FDP 
                 P 
                 2 
                 2.644 
                 73.782 
                 92.229 
                 1.00 
                 62.62 
                 P 
                 C 
               
               
                 ATOM 
                 46473 
                 O1 
                 FDP 
                 P 
                 2 
                 2.081 
                 74.192 
                 93.454 
                 1.00 
                 63.77 
                 P 
                 O 
               
               
                 ATOM 
                 46474 
                 O5 
                 FDP 
                 P 
                 2 
                 4.658 
                 75.066 
                 91.827 
                 1.00 
                 59.94 
                 P 
                 O 
               
               
                 ATOM 
                 46475 
                 C3 
                 FDP 
                 P 
                 2 
                 3.448 
                 74.488 
                 89.952 
                 1.00 
                 61.69 
                 P 
                 C 
               
               
                 ATOM 
                 46476 
                 O3 
                 FDP 
                 P 
                 2 
                 3.791 
                 75.620 
                 89.151 
                 1.00 
                 61.88 
                 P 
                 O 
               
               
                 ATOM 
                 46477 
                 C4 
                 FDP 
                 P 
                 2 
                 4.651 
                 73.547 
                 90.034 
                 1.00 
                 60.93 
                 P 
                 C 
               
               
                 ATOM 
                 46478 
                 O4 
                 FDP 
                 P 
                 2 
                 4.218 
                 72.190 
                 89.924 
                 1.00 
                 62.00 
                 P 
                 O 
               
               
                 ATOM 
                 46479 
                 C5 
                 FDP 
                 P 
                 2 
                 5.236 
                 73.816 
                 91.421 
                 1.00 
                 59.69 
                 P 
                 C 
               
               
                 ATOM 
                 46480 
                 C6 
                 FDP 
                 P 
                 2 
                 6.762 
                 73.833 
                 91.360 
                 1.00 
                 61.28 
                 P 
                 C 
               
               
                 ATOM 
                 46481 
                 O6 
                 FDP 
                 P 
                 2 
                 7.338 
                 74.231 
                 92.605 
                 1.00 
                 62.38 
                 P 
                 O 
               
               
                 ATOM 
                 46482 
                 P2 
                 FDP 
                 P 
                 2 
                 8.939 
                 74.344 
                 92.725 
                 1.00 
                 62.47 
                 P 
                 P 
               
               
                 ATOM 
                 46483 
                 O5P 
                 FDP 
                 P 
                 2 
                 9.300 
                 73.929 
                 94.237 
                 1.00 
                 62.98 
                 P 
                 O 
               
               
                 ATOM 
                 46484 
                 O6P 
                 FDP 
                 P 
                 2 
                 9.527 
                 73.192 
                 91.766 
                 1.00 
                 62.87 
                 P 
                 O 
               
               
                 ATOM 
                 46485 
                 O4P 
                 FDP 
                 P 
                 2 
                 9.438 
                 75.687 
                 92.356 
                 1.00 
                 62.71 
                 P 
                 O 
               
               
                 END