bluuebunny/crossref_metadata_2025 huggingface dataset:
Bovine kidney 3-methylcrotonyl-CoA and propionyl-CoA carboxylases: Each enzyme contains nonidentical subunits
Authors: Lau Edward P. , Cochran Bruce C. , Munson Lianna , Fall R. Ray
Published: May 2006

3-Methylcrotonyl-CoA carboxylase (MCase; EC 6.4.1.4) and propionyl-CoA carboxylase (PCase; EC 6.4.1.3) have been obtained in highly purified form from bovine kidney mitochondria. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that each enzyme is composed of nonidentical subunits, including a smaller biotin-free subunit (
M
r
62,000 and 58,000 for MCase and PCase, respectively), and a larger biotin-containing subunit (
M
r
80,000 and 74,000 for MCase and PCase, respectively). The possibility that these subunits were derived from a single, larger precursor polypeptide via proteolysis was explored by purification and electrophoresis of each enzyme in the presence of protease inhibitors, but no evidence for proteolysis was obtained. Specific antisera directed towards each enzyme were prepared. The anti-PCase preparation was used to precipitate crossreacting PCase from a pig heart extract. Analysis of the immunoprecipitate obtained revealed a biotin-containing polypeptide (
M
r
78,000) and a biotin-free polypeptide (
M
r
55,000), suggesting that pig heart PCase also contains nonidentical subunits analogous to those seen in the kidney mitochondrial MCase and PCase. A bipartite subunit structure may be a common feature in mammalian MCase and PCase.
X-ray diffraction studies of fibers and crystals of deoxygenated sickle cell hemoglobin.
Authors: Magdoff-Fairchild B , Chiu C C
Published: May 2006

Paracrystalline fibers of deoxygenated sickle hemoglobin in erythrocytes or concentrated solutions exhibit a phase transformation to a fully crystalline state. X-ray diffraction patterns of the fiber and crystallites are similar except in two respects: the equatorial spacings of the fibers suggest that they pack into a square lattice with a = 220 A, whereas those of the crystals can be indexed on the basis of a net of 187 A by 54 A, and the second-order near-meridional reflections are strong on the fiber pattern but weak on that of the crystallites. The crystallites are isomorphous with single crystals grown in polyethylene glycol solution at pH 4.5 whole structure has been determined at near-atomic resolution (Wishner, B.C., Ward, K.B. Lattmen, E.E. & Lowve, W.E. (1975) J. Mol. Biol. 98, 179-194). Double filaments of molecules with an axial repeat of 64 A comprise the basic unit of both the crystal and fiber structures. Each filament of the pair is translated with respect to its neighbor by half a molecular diameter along the fiber axis. The two filaments are held together by contacts made by Val 6beta in the molecules of one strand with hydrophobic side chains of the molecule in the neighboring strand. This interaction is probably the cause of the aggregation of filaments into fibers that leads to the sickling of erythrocytes.
Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition
Authors: Goldstein Joseph L. , Ho Y. K. , Basu Sandip K. , Brown Michael S.
Published: May 2006

Resident mouse peritoneal macrophages were shown to take up and degrade acetylated
125
I-labeled low density lipoprotein (
125
I-acetyl-LDL)
in vitro
at rates that were 20-fold greater than those for the uptake and degradation of
125
I-LDL. The uptake of
125
I-acetyl-LDL and its subsequent degradation in lysosomes were attributable to a high-affinity, trypsin-sensitive, surface binding site that recognized acetyl-LDL but not native LDL. When
125
I-acetyl-LDL was bound to this site at 4°C and the macrophages were subsequently warmed to 37°C, 75% of the cell-bound radioactivity was degraded to mono[
125
I]iodotyrosine within 1 hr. The macrophage binding site also recognized maleylated LDL, maleylated albumin, and two sulfated polysaccharides (fucoidin and dextran sulfate) indicating that negative charges were important in the binding reaction. A similar binding site was present on rat peritoneal macrophages, guinea pig Kupffer cells, and cultured human monocytes but not on human lymphocytes or fibroblasts, mouse L cells or Y-1 adrenal cells, or Chinese hamster ovary cells. Uptake and degradation of acetyl-LDL via this binding site stimulated cholesterol esterification 100-fold and produced a 38-fold increase in the cellular content of cholesterol in mouse peritoneal macrophages. Although the physiologic significance, if any, of this macrophage uptake mechanism is not yet known, we hypothesize that it may mediate the degradation of denatured LDL in the body and thus serve as a “backup” mechanism for the previously described receptor-mediated degradation of native LDL that occurs in parenchymal cells. Such a scavenger pathway might account for the widespread deposition of LDL-derived cholesteryl esters in macrophages of patients with familial hypercholesterolemia in whom the parenchymal cell pathway for LDL degradation is blocked, owing to a genetic deficiency of receptors for native LDL.
Identification of a non-H-2 gene (Rfv-3) influencing recovery from viremia and leukemia induced by Friend virus complex.
Authors: Chesebro B , Wehrly K
Published: May 2006

The dominant C57BL/10 allele of a single autosomal, non-H-2 gene (Rfv-3) was found to be required for recovery from viremia and leukemia induced by Friend virus complex in H-2b/b mice. In H-2a/a mice, the Rfv-3 gene apparently influenced recovery from viremia in the presence of persistent leukemia because these mice lacked the appropriate H-2 genotype for recovery from leukemia. The Rfv-3 gene was distinct from the Fv-2 gene because recovery from viremia was seen in recombinant-inbred mice with the Fv-2s/s genotype. Furthermore, backcross studies indicated that Rfv-3 and Fv-2 were not linked. The Rfv-3 gene may act by influencing the specific anti-FV humoral antibody response.
Competitive inhibition of colchicine binding to tubulin by microtubule-associated proteins.
Authors: Nunez J , Fellous A , Francon J , Lennon A M
Published: May 2006

Microtubule-associated proteins (MAPs) promote tubulin polymerization, whereas colchicine inhibits this process. In this paper, MAPs have been shown to inhibit colchicine binding to tubulin in a competitive manner. Attempts were made to identify which of the MAPs fraction(s) was responsible; both tau protein (a thermostable molecule with a molecular weight of approximately 70,000) and a high molecular weight fraction (HMW) were able to compete with colchicine. In contrast, Mg2+, which also induces microtubule assembly in vitro, had no effect on colchicine binding to tubulin.
Structure of the human G gamma-A gamma-delta-beta-globin gene locus.
Authors: Bernards R , Little P F , Annison G , Williamson R , Flavell R A
Published: May 2006

We have constructed a physical map of the human G gamma-, A gamma-, delta-, and beta-globin genes. The previously described maps of the fetal and adult beta-like globin genes have been linked to one another by identification of a DNA fragment, generated by BamHI, that contains part of each of the A gamma- and delta-globin genes. The map obtained, which spans more than 40 kilobases, shows the following intergene distances: between G gamma and A gamma, 3500 base pairs; between A gamma and delta, 13,500 base pairs; and between delta and beta, 5500 base pairs. All genes are transcribed from the same DNA strand.
13C NMR quantitation of polymer in deoxyhemoglobin S gels.
Authors: Noguchi C T , Torchia D A , Schechter A N
Published: May 2006

13C/1H magnetic double-resonance spectroscopy has been used to quantitate the amount of polymerized hemoglobin S in deoxygenated gels at 30 degrees C, for samples whose hemoglobin concentration range from 21 to 32 g/dl. Scalar- and dipolar-decoupled spectra and a 13C proton-enhanced dipolar-decoupled spectrum were recorded for each sample as was a scalar-decoupled spectrum for a matching oxyhemoglobin S control. The difference between the oxyhemoglobin S and deoxyhemoglobin S scalar-decoupled spectra was used to determine the polymer fraction, and this value was compared with the polymer fraction determined by using ultracentrifugation sedimentation on the same sample (assuming a two-phase model). The polymer fraction value determined by uncorrected sedimentation averaged 0.15 more than the value obtained from NMR. The discrepancy between the two techniques was largely removed when the analysis of the sedimentation data included a correction for depletion of hemoglobin in the supernatant or sol phase due to sedimentation of free molecules. The best fit to both the sedimentation and NMR data was obtained by using a solubility of deoxyhemoglobin S at 30 degrees C of 17.3 +/- 1 g/dl. These results indicate that the NMR techniques, which do not require separation of the sample into a sol phase and a pellet phase, provide quantitative information about the deoxyhemoglobin S polymer and will be useful for studies of sickle erythrocytes.
Amino acid sequence of bacteriorhodopsin.
Authors: Khorana H G , Gerber G E , Herlihy W C , Gray C P , Anderegg R J , Nihei K , Biemann K
Published: May 2006

The complete primary structure of the purple membrane protein bacteriorhodopsin, which contains 248 amino acid residues, has been determined. Methods used for separation of the hydrophobic fragments included gel permeation and reverse-phase high-pressure liquid chromatography in organic solvents. The amino acid sequence was determined by a combination of automatic Edman degradation and mass spectrometric methods. The total sequence was derived by ordering of the CNBr fragments on the basis of methionine-containing peptides identified by gas chromatographic mass spectrometry and by analysis of N-bromosuccinimide fragments containing overlaps between CNBr fragments. The present sequence differs from that recently reported by Ovchinnikov and coworkers with respect to an additional tryptophan (position 138) and several amino acid assignments.
On the origin of the contractile force in skeletal muscle.
Authors: Harrington W F
Published: May 2006

Analysis of the early tension responses after abrupt step changes in the length of isometrically contracting skeletal muscle shows that the magnitude of the recovery tension (T2) and the time dependence of this process at various step displacements give a good correlation with the behavior expected for a helix-coil transition in the subfragment-2 (S-2) region of myosin. The "instantaneous" tension response (to T1) after the step change in length appears to have its origin in compliance within the coil region of S-2, which is formed through helix melting at the moment of force generation.
Concomitant synthesis of beta-endorphin and alpha-melanotropin from two forms of pro-opiomelanocortin in the rat pars intermedia.
Authors: Crine P , Gossard F , Seidah N G , Blanchette L , Lis M , Chrétien M
Published: May 2006

In the pars intermedia of rat pituitary glands, two forms of a common precursor for corticotropin (ACTH) and beta-lipotropin with apparent molecular weights of 34,000 and 36,000 were resolved by sodium dodecyl sulfate/acrylamide gradient slab gel electrophoresis. High-performance liquid chromatographic analysis of [35S]methionine-labeled tryptic fragments of the two forms of the precursor revealed that both contained copies of ACTH-(1-8) and beta-lipotropin-(61-69) sequences. When biosynthetic studies were performed in the presence of tunicamycin, the 34,000- and 36,000-dalton forms were replaced by a peptide with an apparent molecular weight of 32,000. It was therefore concluded that the 34,000- and 36,000-dalton forms of the precursor represent two glycoprotein variants of similar polypeptides, differing in the number of asparagine-linked carbohydrate moieties. During pulse-chase incubations with [35S]methionine, the precursor forms were cleaved into two major groups of labeled products: (i) beta-endorphin and (ii) a mixture of ACTH fragments closely related to alpha-melanotropin. No ACTH-(1-39) was found at the end of a 2-hr chase period, suggesting that ACTH is not a significant hormone product of the rat pars intermedia.
Homologous pairing in genetic recombination: complexes of recA protein and DNA.
Authors: Shibata T , Cunningham R P , DasGupta C , Radding C M
Published: May 2006

recA protein, which is essential for general genetic recombination in Escherichia coli, promotes the homologous pairing of single-stranded DNA with double-stranded DNA to form a D loop. The amount of recA protein required for the reaction was directly proportional to the amount of single stranded DNA and was unaffected by similar variations in the amount of double-stranded DNA. The ATP analog, adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), which was not rapidly hydrolyzed by recA protein, blocked the formation of D loops but promoted the formation of stable complexes of recA protein and single-stranded DNA. These complexes, in turn, bound homologous or heterologous double-stranded DNA and partially unwound it. Because ATP gamma S competitively inhibited the ATPase activity of recA protein (Km/Ki approximately 300), we infer that ATP gamma S binds at a site that overlaps the site for ATP and that the functional complexes formed in the presence of the analog probably represent partial steps in the overall reaction. If the complexes formed in the presence of ATP gamma S reflect natural intermediates in the formation of D loops, recA protein must promote homologous pairing either by moving juxtaposed single-stranded and double-stranded DNA relative to one another or by forming and dissociating complexes reiteratively until a homologous match occurs.
In vitro
activation of insect prothoracic glands by the prothoracicotropic hormone
Authors: Bollenbacher Walter E. , Agui Noriaki , Granger Noelle A. , Gilbert Lawrence I.
Published: May 2006

An
in vitro
assay for the prothoracicotropic hormone has been developed that utilizes an ecdysone radioimmunoassay to quantify the increase in the rate of ecdysone synthesis elicited by the neurohormonal activation of the prothoracic glands. The rapidity, reproducibility, and accuracy of the assay were maximized by using one member of a gland pair as the control and the other as the test gland. This was possible because the basal rates of ecdysone synthesis by the members of a gland pair were equivalent. Activation was demonstrated to be dose dependent and specific, with prothoracicotropic hormone activity present only in homogenates of brain. The
in vitro
activation of the prothoracic glands was verified with the
Manduca
bioassay for the prothoracicotropic hormone in which the morphological responses to the hormone were correlated with increased
in vivo
ecdysone titers. These results provide unequivocal evidence that the activation of the prothoracic glands by the prothoracicotropic hormone is direct and suggest that activation represents an increase in a basal rate of ecdysone synthesis.
C4a: the third anaphylatoxin of the human complement system.
Authors: Gorski J P , Hugli T E , Müller-Eberhard H J
Published: May 2006

The activation peptide C4a was isolated from C1s-cleaved C4, the fourth component of complement. The peptide appeared to be homogeneous by electrophoresis on cellulose acetate and by polyacrylamide gel electrophoresis. C4a has a molecular weight of 8650 and an electrophoretic mobility at pH 8.6 of +2.1 x 10(-5) cm2V-1 sec-1. Carboxypeptidase B released approximately 1 mol of arginine per mol of C4a. The partial COOH-terminal sequence was determined to be Leu-Gln-Arg-COOH. The isolated C4a was spasmogenic for guinea pig ileum at a concentration of 1 microM and it desensitized the muscle (i.e., produced tachyphylaxis) with respect to human C3a anaphylatoxin (at 0.33 microM) but not with respect to human C5a anaphylatoxin. Increased vascular permeability was observed in human skin after intradermal injection of 1 nmol of C4a, as evidenced by immediate erythema and edema formation. The spasmogenic, tachyphylactic, and vascular activities of C4a were abrogated by removal of the COOH-terminal arginine, a property that is characteristic also of the C3a and C5a anaphylatoxins. Contamination of C4a with either C3a or C5a has been ruled out by using radioimmunoassays for these peptides. Although C4a is considerably less active than are C3a and C5a, the present observations suggest that C4a constitutes a heretofore unrecognized anaphylatoxin that is related biologically and chemically to the activation peptides of C3 and C5.
Guanine nucleotide-binding activity as an assay for
src
protein of rat-derived murine sarcoma viruses
Authors: Scolnick Edward M. , Papageorge Alex G. , Shih Thomas Y.
Published: May 2006

We have recently identified a 21,000-dalton protein, p21, coded for by Kirsten or Harvey murine sarcoma virus. On the basis of the results obtained with the p21 of a mutant of Kirsten sarcoma virus, temperature sensitive for the maintenance of transformation, we concluded that the p21 was required for the maintenance of transformation induced by either virus. We report herein that when extracts from cells transformed by Kirsten or Harvey sarcoma virus are incubated with [
3
H]GDP or [α-
32
P]GTP, picomole quantities of guanine nucleotide can be immunoprecipitated with antisera that contain antibodies to the p21. Previously we have shown that the immunoprecipitability of [
35
S]methionine-labeled p21 of the temperature-sensitive mutant of Kirsten sarcoma virus is thermolabile. The binding of guanine nucleotide is shown herein also to be thermolabile in extracts of cells transformed by the same mutant. However, the immunoprecipitability of the [
35
S]methionine-labeled p21 in such extracts of the temperature-sensitive mutant can be preserved if the extracts containing labeled p21 are incubated with added GDP or GTP prior to heating. The results suggest an interaction between p21 and certain guanine nucleotides, and the possible roles of guanine nucleotides and p21 in the maintenance of transformation are discussed.
Implantation of the isolated human erythrocyte anion channel into plasma membranes of Friend erythroleukemic cells by use of Sendai virus envelopes.
Authors: Volsky D J , Cabantchik Z I , Beigel M , Loyter A
Published: May 2006

Sendai virus envelopes and human erythrocyte band 3 membrane polypeptides were isolated in detergent solutions and coreconstituted into detergent-free vesicle structurally resembling viral envelopes. Formation of hybrid viral envelope-band 3 vesicles (VB3) was demonstrated by immunoprecipitation of VB3 containing 4,4'-diisothiocyano-2,2'-ditritostilbenedisulfonic acid-labeled band 3 in the presence of antiviral antiserum. The hybrid VB3 vesicles displayed agglutination, fusion, and hemolytic properties similar to those of reconstituted viral envelopes. Incubation of VB3 with Friend erythroleukemic cells resulted in fusion-mediated insertion of viral antigens and of band 3 into the plasma membranes of the recipient cells. The presence of band 3 incorporated into Friend erythroleukemic cells was revealed structurally by electron microscopy after staining with ferritin-conjugated anti-human erythrocyte membrane antiserum. Incorporation of band 3, the putative erythrocyte antion channel, led to a marked and specific stimulation of anion exchange in Friend erythroleukemic cells. The viral-mediated insertion of an isolated membrane protein into viable Friend erythroleukemic cells provides a method for studying membrane protein turnover and for assaying reconstituted membrane components such as carriers or channels.
Distinctive nucleotide sequences of promoters recognized by RNA polymerase containing a phage-coded "sigma-like" protein.
Authors: Talkington C , Pero J
Published: May 2006

We report the nucleotide sequences of two promoters for bacteriophage SP01 "middle" genes. These promoters are recognized by a modified form of Bacillus subtilis RNA polymerase that contains a phage-coded "sigma-like" regulatory protein (gp28) in place of the bacterial sigma factor. Both promoters shared the identical hexanucleotide 5'A-G-G-A-G-A at about 35 base pairs preceding the start point of transcription and the identical heptanucleotide 5'-T-T-T-A-T-T-T (T is the thymine analog 5-hydroxymethyluracil in SP01 DNA) located about 10 base pairs preceding the transcriptional start point. The significance of these sequences in comparison with nucleotide sequences of promoters recognized by sigma-containing RNA polymerases is discussed.
Glycoproteins of Sendai virus are transmembrane proteins.
Authors: Lyles D S
Published: May 2006

Radiolabeled Sendai viral envelope proteins were incorporated into human erythrocyte membranes by the process of fusion of the viral envelope with the erythrocyte membrane. Inside-out (IO) vesicles were prepared from the erythrocyte membranes containing viral proteins, and the presence of viral proteins was assessed by electrophoresis in sodium dodecyl sulfate/polyacrylamide gels. Proteolysis with trypsin, chymotrypsin, or Pronase, which digests only the external surface of the IO vesicles (the cytoplasmic surface of the erythrocyte membrane) revealed that the viral nucleocapsid and the nonglycosylated inner-envelope (M) proteins were present on the external surface. In addition, small segments of the viral envelope glycoproteins (HN and F1) were removed by these proteases, while the major portions of the glycoproteins were protected from digestion, indicating that in the erythrocyte membrane they are transmembrane proteins. Results of experiments carried out on right side-out membranes, unsealed membranes, and membranes containing attached virus that was unable to fuse indicated that the results obtained with IO membranes could not be accounted for by contamination with these membrane species. The identify of the proteolysis products was confirmed by peptide mapping. The selective exposure of the cytoplasmic surface, and hence the internal components of the virus, in IO vesicles makes this membrane system an attractive model for studying the interactions involved in virus maturation at host cell membranes.
Cellular localization of the insect prothoracicotropic hormone:
In vitro
assay of a single neurosecretory cell
Authors: Agui Noriaki , Granger Noelle A. , Gilbert Lawrence I. , Bollenbacher Walter E.
Published: May 2006

The distribution of prothoracicotropic hormone in the pupal brain of
Manduca sexta
has been determined by an
in vitro
assay for prothoracic gland activation. Prothoracicotropic activity was observed in both the brain and retrocerebral complex, but predominantly in the dorosolateral regions of the protocerebrum. Of the two groups of neurosecretory cells present in this area of the brain, only the two lateral type III neurosecretory cells exhibited significant prothoracicotropic hormone activity. Further analysis revealed that the neurohormone was localized in only one of the two type III cells, suggesting that a single neurosecretory cell in each hemisphere is the source of the hormone at the stage examined (day 0). Prothoracicotropic hormone activity was detected in both the corpora allata and the corpora cardiaca, but the corpora allata contained 2 to 9 times the activity of the corpora cardiaca, depending on developmental stage. The significantly higher level of activity in the corpora allata suggests that they may be the neurohemal organs through which the prothoracicotropic hormone of
Manduca
is released.
Changes in plasma lipoprotein distribution and formation of two unusual particles after heparin-induced lipolysis in hypertriglyceridemic subjects.
Authors: Forte T M , Krauss R M , Lindgren F T , Nichols A V
Published: May 2006

Lipoprotein morphology and distribution were studied in three moderately to severely hypertriglyceridemic patients after heparin-induced lipolysis. Lipoproteins with a flotation rate of Sf degrees 12-20 in rho 1.063-g/ml NaCl solution increased substantially in mass 2.5-10 min after heparin administration. This fraction contained 40- to 120-nm flattened particles and increased concentrations of phospholipid and free cholesterol. At 2.5 and 10 min after heparin, the high density lipoproteins (HDL) of rho, 1.126-1.21 g/ml contained small (5.8-nm diameter) spherical particles. Both types of particles disappeared 30-60 min after heparin. Results indicated that with lipolysis HDL3 may be transformed into HDL2a by incorporation of chylomicron and very low density lipoprotein constituents. It is suggested that the 40- to 120-nm particles represent surface fragments of chylomicrons and very low density lipoproteins generated during lipolysis, whereas the 5.8-nm particles are produced either by fragmentation of the large surface constituents or by loss of lipid from existing HDL3.
Galactosamine-induced sensitization to the lethal effects of endotoxin.
Authors: Galanos C , Freudenberg M A , Reutter W
Published: May 2006

Treatment of rabbits, rats, and mice with D-galactosamine increased their sensitivity to the lethal effects of lipopolysaccharide several thousand fold. The susceptibility of the animals was highest when the lipopolysaccharide was injected together with galactosamine and decreased successively when injection was carried out 1, 2, and 3 hr later. Sensitization was absent when the lipopolysaccharide was administered 1 hr before or 4 hr after galactosamine. The onset of lethality after treatment with galactosamine and lipopolysaccharide occurred faster than with lipopolysaccharide alone; usually all animals died 5-9 hr later. The galactosamine-induced sensitization to lipopolysaccharide could be reversed by uridine which is known to inhibit the early biochemical alterations induced by the amino sugar in the hepatocytes. Although galactosamine is known to exhibit hepatotoxic activity inducing ultimate necrosis of the hepatocytes, the data so far suggests that the sensitization to lipopolysaccharide is related only to the early metabolic effects of the hexosamine.
Migration of Schwann cells and wrapping of neurites in vitro: a function of protease activity (plasmin) in the growth medium.
Authors: Kalderon N
Published: May 2006

In vitro conditions were defined under which Schwann cells, from a population of dissociated embryonic chicken spinal cord cells, migrate along the growing neuronal fibers and wrap bundles as well as individual axons, in a pattern similar to that found in a developing peripheral nervous system in vivo. The migration of Schwann cells and their wrapping of nerve fibers was found to be a function of plasmin activity in the growth medium. It was determined that at least one cell type among the spinal cord cells is producing plasminogen activator, the enzyme that activates the plasminogen that is a constituent of any serum. It is concluded that, to achieve wrapping of neurons by Schwann cells in culture, it is essential to have an active plasmin-generating system in the medium. It is hypothesized that the Schwann cell produces plasminogen activator. The possible role of both the Schwann cell and the plasminogen possible role of both the Schwann cell and the plasminogen activator in the formation of the neuromuscular junction is discussed.
Direct spectroscopic observation of singlet oxygen emission at 1268 nm excited by sensitizing dyes of biological interest in liquid solution
Authors: Khan A. U. , Kasha M.
Published: May 2006

The direct observation of dye-photosensitized 1268-nm emission of the
1
Δ
g
→
3
Σ
g
transition of molecular oxygen in liquid solution at room temperature is reported. Singlet oxygen was photosensitized by UV excitation of perfluorobenzophenone in fluorocarbon solvent, by 3,4-benzpyrene and hematoporphyrin in carbon tetrachloride, and by methylene blue in water. Also reported is the development of an extremely sensitive near-infrared spectrophotometer that uses a thermoelectrically cooled lead sulfide detector, optimized optics, and a boxcar integrator as a data processor.
Universal variational functionals of electron densities, first-order density matrices, and natural spin-orbitals and solution of the
v
-representability problem
Authors: Levy Mel
Published: May 2006

Universal variational functionals of densities, first-order density matrices, and natural spin-orbitals are explicitly displayed for variational calculations of ground states of interacting electrons in atoms, molecules, and solids. In all cases, the functionals search for constrained minima. In particular, following Percus [Formula: see text] is identified as the universal functional of Hohenberg and Kohn for the sum of the kinetic and electron—electron repulsion energies of an
N
-representable trial electron density ρ.
Q
[ρ] searches all antisymmetric wavefunctions Ψ
ρ
which yield the fixed. ρ.
Q
[ρ] then delivers that expectation value which is a minimum. Similarly, [Formula: see text] is shown to be the universal functional for the electron—electron repulsion energy of an
N
-representable trial first-order density matrix γ, where the actual external potential may be nonlocal as well as local. These universal functions do not require that a trial function for a variational calculation be associated with a ground state of some external potential. Thus, the
v
-representability problem, which is especially severe for trial first-order density matrices, has been solved. Universal variational functionals in Hartree—Fock and other restricted wavefunction theories are also presented. Finally, natural spin-orbital functional theory is compared with traditional orbital formulations in density functional theory.
Evidence that the avian sarcoma virus transforming gene product is a cyclic AMP-independent protein kinase.
Authors: Erikson R L , Collett M S , Erikson E , Purchio A F
Published: May 2006

The avian sarcoma virus transforming gene product pp60src has been partially purified by using ion exchange or immunoaffinity chromatography. These preparations contain a cyclic AMP-independent protein kinase activity capable of transferring radiolabel from [gamma-32P]ATP to immune rabbit IgG, casein, and the heavy chain purified from normal rabbit IgG. The casein kinase activity is specifically inhibited by anti-pp60src IgG. Comparison of the pp60src-protein kinase isolated from cells transformed by a wild-type ASV or by an ASV temperature-sensitive transformation mutant revealed that the latter product had a half-life of 3 min at 41 degrees C whereas that of the wild-type product was 20 min.
Attractants and repellents influence methylation and demethylation of methyl-accepting chemotaxis proteins in an extract of Escherichia coli.
Authors: Kleene S J , Hobson A C , Adler J
Published: May 2006

During bacterial chemotaxis, attractants and repellents alter the methylation levels of the methyl-accepting chemotaxis proteins (MCPs). These methylation levels represent a balance between two enzymatic processes: methylation and demethylation. In vivo experiments previously have shown that chemoeffectors influence the demethylation process; effects on the methylation system have not been reported. Here we show that in a cell-free extract of Escherichia coli both methylation and demethylation of the MCPs are affected by attractants and repellents. Attractants enhance methylation and inhibit demethylation. Repellents inhibit methylation and stimulate demethylation. The cell-free system provides an opportunity for further study of the mechanisms by which attractants and repellents influence the levels of methylation of the MCPs.
Single-strand binding protein enhances fidelity of DNA synthesis in vitro.
Authors: Kunkel T A , Meyer R R , Loeb L A
Published: May 2006

The effect of Escherichia coli single-strand binding protein on the accuracy of in vitro DNA synthesis has been determined by using two independent methods. By using the synthetic polynucleotide poly[d(A-T)] and measuring dGTP misincorporation or by using phi X174 DNA and measuring nucleotide substitutions, we found that binding protein increases the fidelity of DNA synthesis by as much as 10-fold. This increase is observed with DNA polymerases of divergent sources and is progressive with increasing concentration of binding protein. The increased accuracy observed with DNA polymerases lacking a 3' leads to 5' exonuclease points to a mechanism other than augmented proofreading. In accord with the properties of single-strand binding proteins, it is suggested that increased fidelity is a result of enhanced base selection by the DNA polymerase, resulting from increased rigidity of the template due to its interaction with binding protein.
Nucleosome organization of the yeast 2-micrometer DNA plasmid: a eukaryotic minichromosome.
Authors: Nelson R G , Fangman W L
Published: May 2006

The eukaryotic microorganism Saccharomyces cerevisiae contains 50-100 copies per cell of a circular plasmid called 2-micrometer DNA. The intracellular structure of these molecules, which represent about 4% of the total DNA, was examined by digestion of total cellular chromatin with micrococcal nuclease (nucleate 3'-oligonucleotidohydrolase, EC 3.1.31.1). Nuclease-resistant DNA fragments were fractionated by gel electrophoresis and 2-micrometer DNA sequences were detected by hybridization. The 2-micrometer and chromosomal DNA digestion patterns were very similar indicating that both types of DNA are condensed into nucleosomes. An analysis of these digestion patterns showed that the kinetics of digestion of 2-micrometer chromatin and total chromatin are similar and that both have the same nucleosome repeat length of about 165 base pairs. Native 2-micrometer plasmids were examined by zone sedimentation in sucrose gradients containing 0.15 M NaCl and were found to have a sedimentation constant of 75 S, about 3 times the sedimentation constant of protein-free 2-micrometer DNA. This sedimentation property is what would be expected for a 2-micrometer DNA minichromosome. We conclude that within the cell 2-micrometer DNA molecules are organized in a chromatin structure very similar to that of the yeast chromosomes.
Five TGA “stop” codons occur within the translated sequence of the yeast mitochondrial gene for cytochrome
c
oxidase subunit II
Authors: Fox Thomas D.
Published: May 2006

A mitochondrial mutation that genetically maps in the middle of the gene coding cytochrome
c
oxidase subunit II has been found to be a single-base-pair deletion. Three independently isolated spontaneous revertants of this mutant have different single-base-pair insertions within 15 nucleotides of the mutation. These findings clearly identify the location of the gene and suggest that the mutation causes a frame-shift. The sequence of about 900 base pairs surrounding the mutation has been determined and found to have several chain termination codons in every possible reading frame. The sequence can, however, be translated in one frame by assuming that the codon TGA does not cause chain termination in yeast mitochondira, as was recently suggested for the human organelle [Barrell, B. G., Bankier, A. T. & Drouin, J. (1979)
Nature
(
London
), in press]. If TGA codes for tryptophan residues, as is apparently the case in human mitochondria, a polypeptide can be read from the yeast mtDNA that is identical to bovine cytochrome oxidase subunit II at 37.8% of its residues. Furthermore, the DNA sequences of the frame-shift revertants discussed above predict relative isolectric point differences between the wild-type and various revertant forms of the polypeptide. The detection of these isolectric point differences by two-dimensional electrophoresis of subunit II from the various strains independently confirms the presumed reading frame of the gene. It is concluded that TGA is translated in yeast mitochondria, most probably as tryptophan.
Enhancement of certain biological activities of muramyl dipeptide derivatives after conjugation to a multi-poly(DL-alanine)--poly(L-lysine) carrier.
Authors: Chedid L , Parant M , Parant F , Audibert F , Lefrancier F , Choay J , Sela M
Published: May 2006

N-Acetylmuramyl-L-Ala-D-Glu-NH2 (muramyl dipeptide) and several of its derivatives are effective immunoactivators that can enhance nonspecific resistance to infection but can also elicit fever. In contrast, one of its stereoisomers, N-acetylmuramyl-D-Ala-D-Glu-NH2, is devoid of both these activities. Our present report demonstrates that macromolecularization of muramyl dipeptide by attachment of several units to a multi-poly(DL-Ala)-poly(L-Lys) carrier potentiates both its pyrogenic and its immunostimulant activity. This branched polymer has been extensively used as carrier to various haptens. Surprisingly, inactive N-acetylmuramyl-D-Ala-D-Glu-NH2, after conjugation under the same conditions, becomes capable of increasing nonspecific immunity although its lack of pyrogenicity is not greatly modified. Moreover, the N-acetylmuramyl-D-Ala-D-Glu--NH2 conjugate remains devoid of adjuvant, sensitizing, or eliciting activity.
Gravitational interaction of hadrons and leptons: Linear (multiplicity-free) bandor and nonlinear spinor unitary irreducible representations of
S̄L̄
(4
R
)
Authors: Ne'eman Yuval , Šijački Djordje
Published: May 2006

We review two possible affine extensions of gravity connected to the strong interactions. In the metric affine theory, torsion and nonmetricity do not propagate, gravitation is effectively unmodified, and the observed approximate conservation of hadron intrinsic hypermomentum—i.e., scaling,
SU
(6), and Regge trajectories—is due to the
ḠL̄
(4,
R
) band-spinor structure of the hadrons. In the second approach, the new gravitational Lagrangian density generates propagating but confined torsion and nonmetricity, presumably the main contributions to quark confinement. Leptons are represented nonlinearly as Poincaré spinors with the metric field as “realizer” and Higgs boson, and are unconfined. We present a construction for all linear multiplicity-free (= bandor) representations of
ḠL̄
(4,
R
) and in particular the [Formula: see text] fitting the hadron manifield. We also construct the Hilbert space hadron states [irreps (irreducible representations) of
ḠĀ
(4,
R
)] and the nonlinear realizations of
ḠL̄
(4,
R
) for lepton fields.
Origin and direction of DNA replication of plasmid RSF1030.
Authors: Conrad S E , Wold M , Campbell J L
Published: May 2006

An in vitro replication system has been used to study the origin and direction of replication of the covalently closed, circular DNA of plasmid RSF1030, a nonconjugative R factor. We have enriched for replicative intermediates in these studies either by isolating them on the basis of their unique structure or by limiting the extent of synthesis in the in vitro system. Circular molecules that have replicated to various extents migrate to characteristic positions in agarose gels, thus providing a rapid and efficient method for isolating partially replicated forms. Alternatively, replicative intermediates can be isolated directly from reaction mixtures that contain dideoxyTTP (ddTTP), a compound that limits the average extent of synthesis in vitro. Electron microscopic analysis of such intermediates linearized with either Hpa I or BamHI indicates that RSF1030 replicates in vitro from a unique origin located 70% from one end of Hpa I-cleaved molecules and 47% from the BamHI site. The unidirectional mode of replication has been confirmed by the order in which the six HincII fragments of RSF1030 DNA are labeled in vitro when synthesis is limited to various extents with ddTTP. Finally, a physical map of RSF1030 has been constructed using the restriction endonucleases BamHI, Hpa I, and HincII, and the origin and direction of replication have been defined relative to the map.
Stability of lipid vesicles in tissues of the mouse: a gamma-ray perturbed angular correlation study.
Authors: Mauk M R , Gamble R C
Published: May 2006

The rate of phospholipid vesicle disruption in specific tissues of the mouse was followed by gamma-ray perturbed angular correlation (PAC) spectroscopy. In these studies, high levels of 111In-nitrilotriacetic acid complex are contained in unilamellar vesicles consisting of distearoyl phosphatidylcholine, cholesterol, and small amounts of other lipids which modify the surface properties. The PAC technique monitors the extent of vesicle breakup by measuring a time-integrated perturbation factor, less than G22 (infinity) greater than. As the vesicles are broken open in vivo, the released 111In3+ ions quickly bind to macromolecules and the less than G22 (infinity) greater than value decreases substantially. After administration of vesicles by various routes (intravenous, intraperitoneal, subcutaneous, and oral), the radioactivity and less than G22 (infinity) greater than values were determined for several tissues at intervals up to 24 hr. We conclude from these data that (i) the PAC technique in conjunction with standard gamma counting methods provides unique information on the condition and location of vesicles in specific tissues, (ii) significant differences in vesicle integrity are found in various tissues, and (iii) both the means of administration and the presence of surface charge affect the vesicle stability and distribution. The carbohydrate analogues of cholesterol affect vesicle stability but not distribution.
Desensitization of the insulin receptor at an early postreceptor step by prolonged exposure to antireceptor antibody
Authors: Karlsson F. Anders , Obberghen Emmanuel Van , Grunfeld Carl , Kahn C. Ronald
Published: May 2006

We have used an adipocyte-like cell line, the 3T3-L1 fatty fibroblasts, to compare acute and chronic effects of autoantibodies directed against the insulin receptor. Acute exposure of the cells in tissue culture to the antibodies resulted in a blockade of insulin binding and stimulation of 2-deoxy-glucose transport and glucose oxidation. Maximal acute effects were reached within 30-120 min. Subsequently, the stimulatory response decayed and, after 6 hr in the continuous presence of the antibodies, basal glucose metabolism had returned to the level observed with unexposed cells and a state of severe insulin resistance prevailed. In contrast to the decay of bioresponse, no change in insulin binding was detectable over the same time period. The mechanism of desensitization seemed to involve events early after insulin binding to receptor because cells exposed to antibody for prolonged periods of time, although unresponsive to insulin and antireceptor antibodies, responded normally to both spermine and vitamin K
5
, agents that stimulate glucose metabolism independently of the insulin receptor. These data suggest that prolonged or continuous occupancy of the insulin receptor by a ligand, in this case antireceptor antibodies, does not produce a continuous biological response. Instead, there is desensitization at some early step in the pathway for insulin action. These observations have important implications with respect to the mechanism of insulin action and to other situations in which there is long-term exposure of cells to antibodies that react with membrane components.
Molecular evolution of biomembranes: structural equivalents and phylogenetic precursors of sterols.
Authors: Rohmer M , Bouvier P , Ourisson G
Published: May 2006

Derivatives of one triterpene family, the hopane family, are widely distributed in prokaryotes; they may be localized in membranes, playing there the same role as sterols play in eukaryotes, as a result of their similar size, rigidity, and amphiphilic character. Their biosynthesis embodies many primitive features compared to that of sterols and could have evolved toward the latter once aerobic conditions had been established. Membrane reinforcement appears to be achieved in other prokaryotes by other mechanisms, involving either approximately 40-A-long rigid hydrocarbon chains terminated by one polar group acting like a peg through the double-layer or similar chains terminated by two polar groups acting like tie-bars across the membrane. These inserts can be tetraterpenes (e.g., carotenoids). The biophysical function of membrane optimizers appears to have evolved toward sterols by changes limited to only a few enzymatic steps of the same fundamental biosynthetic processes.
Genetic heterogeneity of "normal" human erythrocyte glucose-6-phosphate dehydrogenase: an isoelectrophoretic polymorphism.
Authors: Modiano G , Battistuzzi G , Esan G J , Testa U , Luzzatto L
Published: May 2006

Quantitative determination of glucose-6-phosphate dehydrogenase (G6PD; D-glucose-6-phosphate: NADP+ 1-oxidoreductase, EC 1.1.1.49) activity was carried out in 214 male Nigerian children of 84 mothers with known Gd genotype. The relative intrasibship difference in G6PD activity (normalized to the lowest value within the sibship) was below 0.18 in all cases but one when the children were known to have the same Gd+ allele (identical by descent); whereas it was higher than 0.18 in 18 out of 33 sibships in which children might have had either of the two maternal (electrophoretically identical) Gd+ alleles. G6PD from 10 (8 G6PD B and 2 G6PD A) children belonging to four of the sibships possessing high quantitative variation in G6PD activity was partially purified and extensively characterized. The 8 G6PD type B samples fell unambiguously into two classes on the basis of Km values for glucose 6-phosphate (determined at variuos pH values), and KCl gradient elution from DEAE-Sephadex columns. The two types of G6PD B were resolved from an artificial mixture on a DEAE-Sephacel column. The two G6PD type A samples were also different from each other by the same criteria. We conclude that "normal" G6PD is genetically heterogeneous and that the structural Gd alleles concerned are all polymorphic in the Nigerian population. In this instance, a human enzyme polymorphism, not associated with enzyme deficiency, is revealed by an approach other than electrophoresis.
Regulation of macrophage migration by products of the complement system.
Authors: Bianco C , Götze O , Cohn Z A
Published: May 2006

Agents formerly shown to induce rapid macrophage spreading were examined for their ability to modify the migration of macrophages in the capillary tube assay. Products of the activation of the contact phase of blood coagulation as well as the purified component Bb, the large cleavage fragment of factor B of the alternative complement pathway produced a dose-dependent inhibition of migration. In addition, inflammatory macrophages elicited with either a lipopolysaccharide endotoxin or thioglycollate medium exhibited rapid spreading and inhibited migration, whereas resident cells did not. A close correlation existed, therefore, between enhanced spreading and inhibited migration under both in vitro induced and in vivo situations. Cleavage products of component C5 of the classical complement pathway enhanced macrophage migration and did not alter spreading. In mixtures of C5 cleavage products and Bb, the predominant peptide determined the outcome of the reaction. Factor B, a normal secretory product of macrophages, may represent a common substrate for several of the proteases that induce spreading, inhibit migration, and lead to the generation of the enzymatically active fragment Bb.
Regulation of immune response to tumor antigen: interference with syngeneic tumor immunity by anti-IA alloantisera.
Authors: Perry L L , Dorf M E , Benacerraf B , Greene M I
Published: May 2006

We present evidence for a role of I-A subregion-encoded determinants in syngeneic tumor immunity. In animals rendered immune to the S1509a fibrosarcoma, daily treatment with microliter quantities of antisera directed against Kk and I-Ak determinants expressed on lymphoid cells of host origin decreased the capacity for immune tumor rejection. Absorption studies revealed that anti-I-Ak antibody activity alone was sufficient for the manifestation of this effect. Furthermore, experiments utilizing F1 hybrids showed that an antiserum that was genetically unable to interact with H-2 determinants expressed on the tumor was equally effective in inhibiting tumor immunity. Suggestive evidence that the activity of this antiserum is related to interference with the generation of effector T cell function was provided by the observation that hyperimmune animals pretreated with an anti-Kk,I-Ak antiserum were no longer capable of adoptively transferring tumor immunity to naive recipients. Thus, it is possible to regulate the secondary immune response to tumor antigens by using antisera with specificity for I-A determinants expressed on cells or possibly on factors of the host lymphoid system.