bluuebunny/crossref_metadata_2025 huggingface dataset:
Target Cell Polarity and Membrane Phosphorylation in Relation to the Mechanism of Action of Antidiuretic Hormone
Authors: Schwartz Irving L. , Shlatz Linda J. , Kinne-Saffran Evamarie , Kinne Rolf
Published: May 2006

The plasma membrane of the bovine renal collecting duct epithelial cell has been resolved into its apical (luminal) and basal-lateral (contraluminal) components by free flow electrophoresis. The contraluminal, but not the luminal, membrane was found to contain antidiuretic hormone-sensitive adenylate cyclase. The luminal membrane was found to contain a cyclic 3′:5′-adenosine monophosphate-sensitive self-phosphorylating system consisting of a membrane-bound protein kinase and its membrane-bound substrate(s); this intrinsic protein kinase was not present in the contraluminal membrane.

These findings provide direct evidence that the initiating steps in the action of antidiuretic hormone on the kidney take place at the contraluminal pole of the hormonesensitive target cell and that the late or terminal steps occur at the luminal pole, where they involve an alteration in the level of membrane phosphorylation.
Establishment of a Tetraploid, Immunoglobulin-Producing Cell Line from the Hybridization of Two Human Lymphocyte Lines
Authors: Bloom Arthur D. , Nakamura Frank T.
Published: May 2006

We here report the establishment of a seemingly permanent hybrid cell line formed by fusion of the cells of two biochemically mutant human lymphocyte lines. One parental line (UM-1-6TGr) was deficient in hypoxanthine-guanine phosphoribosyl transferase (IMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.8), and had two marker chromosomes. The second parental line (UM-21-5) was a clonal derivative of a citrullinemic lymphocyte line, and was, like the line of origin, dificient in argininosuccinic acid synthetase [
L
-Citrulline:
L
-aspartate ligase (AMP-forming), EC 6.3.4.5]. This line also had a marker chromosome, which was a B5 with a very prominent secondary constriction. After trypsinization of both parental lines, followed by addition to the fusion mixture of
β
-propiolactone-inactivated Sendai virus, the cells were placed in a doubly selective medium (hypoxanthine-aminopterin-thymidine-containing medium in which the arginine was replaced with citrulline) to prevent the proliferation of the mutant parents. Under selective conditions, 97-99% of cells were found to be tetraploid, containing the three marker chromosomes; and the specific activities of the hybrid line transferase and synthetase were intermediate between normal and mutant line values. Furthermore, the UM-1-6TGr and UM-21-5 lines were producers of gamma and mu heavy chains of immunoglobulin, and of kappa light chains, as determined by immunodiffusion and immunofluorescence, and the hybrid line continued to synthesize and to secrete detectable levels of these same immunoglobulins. These studies demonstrate the genic and cytogenetic stability of this hybridized lymphocyte cell line, and prove that hybridization
per se
does not extinguish the activity of either the regulatory of structural genes involved in immunoglobulin synthesis.
The Structure and Synthesis of Malformin A
Authors: Bodanszky Miklos , Stahl Glenn L.
Published: May 2006

A structure (the disulfide form of cyclo-D-cysteinyl-L-valyl-D-cysteinyl-D-leucyl-L-isoleucyl), previously proposed for malformin A, was reexamined. On the basis of chemical degradations, a different structure (the disulfide form of cyclo-D-cysteinyl-D-cysteinyl-L-valyl-D-leucyl-L-isoleucyl) was established. Accordingly, a compound with this structure was synthesized and was found to be identical with malformin A. The synthetic product causes curvatures on corn roots; maximum effect was seen at a concentration of 0.1 μg/ml, the optimal concentration for malformin A.
Nuclear Magnetic Resonance Spectroscopy. Ring-Current Effects on Carbon-13 Chemical Shifts
Authors: Vernet Richard Du , Boekelheide V.
Published: May 2006

A comparison of the
13
C nuclear magnetic resonance chemical shifts of some 15,16-dialkyl-15,16-dihydropyrenes with the corresponding 15,16-dialkyl-2,7,15,16-tetrahydropyrenes provides a measure of the effect of ring current on carbon-13 chemical shifts in which other effects may be expected to be negligibly small. The general conclusion is that the absolute magnitude of the ring-current effect is the same for carbon-13 as for protons when they occupy the same position in space relative to the aromatic π-electron cloud.
A Calcium-Dependent Mechanism for Synapse and Nerve Cell Membrane Modulation
Authors: Hydén Holger
Published: May 2006

A theory is presented according to which the neuronal membrane and synapses are influenced by membrane-bound S100 protein and a membrane-associated network of actin-like, 20-Å-diameter, coiled filaments and the competition by both proteins for Ca
2+
. The discussion is based on observations made with fluorescein- and peroxidase-conjugated antiserum against S100 protein on isolated nerve cells. A small part of the S100 protein is membrane-bound in a polar localization that develops postnatally. The competition for Ca
2+
can influence the state of the network filaments which uncoil on binding Ca
2+
and affect differently the conformational state of synapses and membrane depending on the S100 protein content.
Kinetics of RNA Labeling in Toad Bladder Epithelium: Effects of Aldosterone and Related Steroids
Authors: Rossier B. C. , Wilce P. A. , Edelman I. S.
Published: May 2006

Sucrose gradient analysis revealed an aldosterone-dependent increase in the incorporation of [
3
H]uridine into a nonmethylated, rapidly labeled RNA that sedimented at 9-12 S. This effect was antagonized by an anti-mineralocorticoid, spironolactone (SC 9420), and was not elicited either by a glucocorticoid, cortisol, or by the inactive isomer, 17-α-isoaldosterone. These findings are in accord with the inference that mRNA is induced by aldosterone during the latent period and that the induced mRNA mediates the action on sodium transport.
Effect of Viscosity on Swimming Velocity of Bacteria
Authors: Keller Joseph B.
Published: May 2006

A theory is given for the swimming velocity of a flagellated bacterium and its variation with the viscosity of the surrounding fluid. The theory is shown to agree fairly well with the experimental observations of Schneider and Doetsch on
Pseudomonas aeruginosa
for viscosities greater than 2 centipoise. However, it fails to account for the decrease of velocity observed when the viscosity decreases from 2 to 1.1 centipoise.
Evidence of Particle-Associated RNA-Directed DNA Polymerase and High Molecular Weight RNA in Human Gastrointestinal and Lung Malignancies
Authors: Cuatico W. , Cho J.-R. , Spiegelman S.
Published: May 2006

Previous communications have demonstrated that neoplastic cells of human breast cancers, leukemias, lymphomas, sarcomas, and brain tumors contain particles with similar diagnostic attributes as those found in RNA oncornaviruses. The present paper concerns malignancies of the gastrointestinal and pulmonary systems for which, like brain tumors, no suitable animal model or corresponding virus exists. By means of the simultaneous detection assay, these tumors have been found to contain 70S RNA and RNA-directed DNA polymerase encapsulated in particulate components possessing densities of 1.16-1.17 g/ml. Twelve out of 17 (70%) colon carcinomas, three out of five (60%) gastric carcinomas, all of three rectal carcinomas, and seven out of ten (70%) lung carcinomas contained detectable levels of these virus-like entities. None of the corresponding normal tissues was positive.
Probability of Gene Fixation in an Expanding Finite Population
Authors: Kimura Motoo , Ohta Tomoko
Published: May 2006

A mathematical theory was developed, based on diffusion models, that enables us to compute the probability of a rare mutant allele eventually spreading through a population when the population size changes with time. In particular, we elaborated the case in which the mutant allele has a definite selective advantage and the population expands following the logistic law. In this case, the probability of ultimate fixation of a single mutant is given by
u
=
2s
(
Z/N
), where
s
is the selective advantage and
Z/N
is a factor by which the probability of fixation is modified through population expansion. Analytical expression was obtained for
Z/N
, and the validity of the formula for
u
was checked by Monte Carlo experiments.
Prostaglandin Synthetase System—Resolution into Oxygenase and Isomerase Components
Authors: Miyamoto Tsumoru , Yamamoto Shozo , Hayaishi Osamu
Published: May 2006

The microsomal fraction of bovine vesicular gland catalyzed the conversion of [
1
-
14
C]8,11,14-eicosatrienoic acid to prostaglandin E
1
in the presence of tryptophan, hemoglobin, and glutathione. The prostaglandin synthetase system was solubilized by treatment of the microsomal fraction with Tween 20 in the presence of ethylene glycol. DEAE-cellulose column chromatography separated the enzyme into two fractions (Fractions I and II), both of which were required for prostaglandin E
1
synthesis. When Fraction I alone was incubated with 8,11,14-eicosatrienoic acid, an unstable compound accumulated. This compound was converted to prostaglandin E
1
by the addition of Fraction II. On the basis of its

R
F

values on thin-layer chromatography, its reduction to prostaglandin F
1α
with stannous chloride, and the decomposition to prostaglandins F
1α
, E
1
, and D
1
at room temperature, the unstable intermediate was tentatively identified to be the 9,11-endoperoxide derivative, referred to previously as prostaglandin R
1
(Nugteren
et al.
(1973)
Biochim. Biophys. Acta
326, 448-461) or prostaglandin H
1
(Hamberg
et al.
(1974)
Proc. Nat. Acad. Sci. USA
71, 345-349). Of the three cofactors mentioned above, Fraction I required both tryptophan and hemoglobin, while only glutathione was necessary for Fraction II. Anti-inflammatory agents such as indomethacin and aspirin were inhibitory to Fraction I.
Pre-proparathyroid Hormone: A Direct Translation Product of Parathyroid Messenger RNA
Authors: Kemper Byron , Habener Joel F. , Mulligan Richard C. , Potts John T. , Rich Alexander
Published: May 2006

An 8-15S RNA fraction from calf parathyroid glands stimulated the incorporation of radioactive lysine and methionine into protein by 15- to 30-fold in a wheat germ extract. The major product, representing 25% of the total protein synthesized, could be bound to an antiserum to parathyroid hormone and binding was inhibited by parathyroid hormone. The chromatographic mobilities of the two tryptic peptides of the cell-free product that contained methionine were identical to the corresponding peptides of parathyroid hormone. Upon electrophoresis in acidic or sodium dodecyl sulfate-acrylamide gels, the cell-free product migrated more slowly than either parathyroid hormone or its biosynthetic precursor, proparathyroid hormone. Analysis of cyanogen bromide products indicated that the cell-free product contained an additional sequence of amino acids at the amino-terminal end. A protein corresponding to the cell-free product could not be detected in intact cells even during incubations with [
3
H]leucine as short as 2 min, which suggests the protein may be a transient precursor to proparathyroid hormone.
Isolation of a polypeptide that has lymphocyte-differentiating properties and is probably represented universally in living cells.
Authors: Goldstein G , Scheid M , Hammerling U , Schlesinger D H , Niall H D , Boyse E A
Published: May 2006

A polypeptide of 8500 molecular weight is described that induces the differentiation of T (thymus-derived) cell and B (bone-marrow-derived) cell immunocytes in vitro, apparently via beta-adrenergic receptors and adenylate cyclase activation. This polypeptide shows a high degree of evolutionary conservation, exhibiting close structural, functional, and immunological similarity when isolated from such diverse origins as cells of mammals and higher plants. This polypeptide was detected in animal cells, yeast, bacteria, and higher plants, and so may well be a universal constituent of living cells.
The antagonistic effect of an inhalation anesthetic and high pressure on the phase diagram of mixed dipalmitoyl-dimyristoylphosphatidylcholine bilayers.
Authors: Trudell J R , Payan D G , Chin J H , Cohen E N
Published: May 2006

Several workers have shown that phase transition-related changes in membrane lipids have a profound effect on membrane-solvated protein function. This phase transition temperature dependence has been explained as resulting from the formation of lateral phase separations within the membrane bilayer. The present study demonstrates that a clinical concentration of an inhalation anesthetic produces changes in both the phase transition temperature of pure lipid bilayers and the lateral phase separation temperature of mixed dipalmitoyl- and dimyristoylphosphatidylcholine bilayers of a magnitude sufficient to influence protein function. It is further shown that pressure is able to antagonize the effect of the anesthetic on these transition temperatures. It is proposed that anesthetic action within nerve membranes may be the result of changes in the lateral phase separation-controlled environment of the membrane-solvated proteins essential to nerve function.
Crystallization and Crystal Data of Monellin
Authors: Wlodawer Alexander , Hodgson Keith O.
Published: May 2006

Crystals of monellin, a sweet protein from
Dioscoreophyllum cumminsii
, were grown by vapor diffusion of 20% ethanol into buffered protein solution. The crystals are orthorhombic, belonging to space group P2
1
2
1
2, with
a
= 54.4 Å,
b
= 113.0 Å,
c
= 40.8 Å, and V = 250,300 Å
3
. The asymmetric unit contains two complete molecules of monellin. The diffraction pattern of this crystal form extends to at least 2.5 Å, indicating that x-ray structural analysis is possible to near-atomic resolution.
Model of protein folding: inclusion of short-, medium-, and long-range interactions.
Authors: Tanaka S , Scheraga H A
Published: May 2006

A hypothesis for protein folding is proposed, in which the native structure is formed by a three-step mechanism: (A) formation of ordered backbone structures by short-range interactions, (B) formation of small contact regions by medium-range interactions, and (C) association of the small contact regions into the native structure by long-range interactions. Empirical interaction parameters (free energy of formation of a contact) between amino-acid residues were evaluated from the frequency of contacts in the x-ray structures of native proteins. On the basis of this mechanism, a Monte Carlo simulation of protein folding (with an accompanying decrease in the total contact free energy) was carried out for bovine pancreatic trypsin inhibitor. The predicted three-dimensional structure is in fairly good agreement with the experimental one.
Catalysis by acetylcholinesterase: evidence that the rate-limiting step for acylation with certain substrates precedes general acid-base catalysis.
Authors: Rosenberry T L
Published: May 2006

Inferences about the catalytic mechanism of acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) are frequently made on the basis of a presumed analogy with chymotrypsin, EC 3.4.21.1. Although both enzymes are serine hydrolases, several differences in the steady-state kinetic properties of the two have been observed. In this report particular attention is focused on the second-order reaction constant, kcat/Kapp. While the reported pH dependence and deuterium oxide isotope effect associated with this parameter for chymotrypsin are generally consistent with simple models involving rate-limiting general acid-base catalysis, this study finds a more complicated situation with acetylcholinesterase. The apparent pKa of kcat/Kapp for acetylcholinesterase varies between 5.5 and 6.3 for neutral substrates and involves nonlinear inhibition by [H+]. Deuterium oxide isotope effects for kcat/Kapp range from 1.1 for acetylcholine to 1.9 for p-nitrophenyl acetate. The bimolecular reaction rate appears rate-limiting for acetylcholine at low concentrations, while a rate-limiting induced-fit step is proposed to account for apparent pKa values and low deuterium oxide isotope effects associated with low concentrations of phenyl acetate and isoamyl acetate.
Oxygen and cyclic nucleotides in human umbilical artery.
Authors: Clyman R I , Blacksin A S , Manganiello V C , Vaughan M
Published: May 2006

In the human umbilical artery O2 has a direct contractile effect and is also required for induction of contraction by several other agents. Agonist that cause contraction (bradykinin, histamine, and serotonin) cause accumulation of guanosine 3':5'-monophosphate (cGMP) without altering adenosine 3':5'-monophosphate (cAMP). They appear to act through two different mechanisms: one Ca++-dependent, the other Ca++-inhibited. O2 increased the cGMP content of the artery in a Ca++-dependent manner without affecting the cAMP content. Inhibitors of oxidative phosphorylation (oligomycin and 2,4-dinitrophenol) did not diminish this effect of O2. O2 was required for demonstration of the Ca++-dependent accumulation of cGMP in response to bradykinin, histamine, and ionophore A23187. The effect of the phosphodiesterase inhibitor 3-isobutyl-1-methyl xanthine on basal cGMP content and on the bradykinin-induced accumulation was also dependent on the presence of O2. Methylene blue and sodium ascorbate caused cGMP accumulation in O2-deprived arteries. Their effects were not diminished in Ca++-depleted arteries and, in fact, seemed to be inhibited when 2.7 mM Ca++ was present in the medium. The effects of these agents and of serotonin on cGMP, which were inhibited by Ca++, were also inhibited by O2. These non Ca++-, non O2-dependent agents (methylene blue, ascorbate, and serotonin) did not, however, permit demonstration of the effects of the Ca++- and O2-dependent agonists on O2-deprived arteries. It appears that there are in the umbilical artery (and probably in other tissues also) at least two separate mechanisms for control of cGMP synthesis that are influenced differently by Ca++- and O2-linked processes.
Yeast manno-protein biosynthesis: solubilization and selective assay of four mannosyltransferases.
Authors: Nakajima T , Ballou C E
Published: May 2006

Using appropriate yeast strains and exogenous acceptors, we have devised specific assays for four mannosyltransferase activities involved in biosynthesis of the carbohydrate outer chain of yeast mannoproteins. The assays utilize GDP-[14C]mannose as the donor and unlabeled oligosaccharides as the acceptors, the products being neutral radioactive oligosaccharides one mannose unit larger than the acceptors. The multiglycosyltransferase system from Saccharomyces cerevisiae was solubilized in Triton X-100 and urea and purified 100-fold. Free mannose is an acceptor for the alpha1 leads to 2-mannosyltransferase, the major product being alpha[14C]Man leads to 2Man. The alpha1 leads to 6-mannooligosaccharides serve as acceptors for both the alpha1 leads to 2- and alpha1 leads to 6-transferases, but the tetrasaccharide alphaMan leads to 3alphsMan leads to 2alphaMan is a specific acceptor for the latter enzyme and yields (see article). When reduced, this same tetrasaccharide serves as the acceptor for an alpha1 leads to 3-mannosyltransferase from Saccharomyces chevalieri, yielding a pentasaccharide with two terminal 1 leads to 3 linkages. Assay of the alpha1 leads to 3-transferase in S. cerevisiae utilizes reduced alpha1 leads to 2-mannotriose as the acceptor, the product being alpha[14C]Man leads to 3alphaMan leads to 2alphaMan leads to 2Mannitol. The multienzyme system works in concert to make "mannan" in a cell-free in vitro system.
Regulation of synthesis of hepatic fatty acid synthetase: binding of fatty acid synthetase antibodies to polysomes.
Authors: Alberts A W , Strauss A W , Hennessy S , Vagelos P R
Published: May 2006

Mammalian fatty acid synthetase was shown to be composed of two peptides, molecular weight 240,000, after dissociation with sodium dodecyl sulfate. Rat liver polysomes that synthesize fatty acid synthetase were identified by sucrose gradient analysis of polysomes that had been reacted with 125I-labeled antibody against fatty acid synthetase. The binding of 125I-labeled antibody to polysomes was found to correlate with the rate of hepatic fatty acid synthesis in various nutritional conditions.
Components of O2-uptake by excised frog nerve dependent upon externally supplied sodium ions.
Authors: Brink F
Published: May 2006

The steady rate of oxygen uptake of excised frog nerves equilibrated in a solution having a very low concentration of sodium ions increases to a new high steady rate when equilibrated with a solution containing a high concentration of this ion. The increase is suppressed by ouabain, indicating participation of the sodium pump. Part of this sodium-activated increase in oxygen uptake is inhibited by tetrodotoxin, indicating that passive influx of sodium ions into axons is part of the total process. Thus, two pathways for passive sodium influx into axons are suggested by these experiments. Procedures known to increase the passive permeability of axons for sodium ions also increase this sodium-activated oxygen uptake. A mechanism is proposed to explain that part of the sodium-activated oxygen uptake that is inhibited by tetrodotoxin.
Effect of oncogenic virus on muscle differentiation.
Authors: Holtzer H , Biehl J , Yeoh G , Meganathan R , Kaji A
Published: May 2006

Chick muscle cultures infected with wild-type Rous sarcoma virus form myotubes, but these myotubes vacuolate and by day 6 most have degenerated, leaving only large numbers of transformed mononucleated, replicating cells. Muscle cultures infected with a temperature-sensitive mutant (TS) at permissive temperatures behave as cells infected with wild-type Rous sarcoma virus. TS-infected cells reared for 8 days at nonpermissive temperature form contracting myotubes, plus large numbers of fibroblastic cells. If these cultures are lowered to permissive temperature, within 72 hr the myotubes vacuolate and degenerate, whereas the mononucleated cells transform. If replicating TS-transformed cells after 8 days at permissive temperature are shifted to nonpermissive temperature, within 72 hr many cells fuse and form contracting, post-mitotic myotubes. Creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) levels parallel the formation and degeneration of myotubes during these temperature shifts. If the viral transforming gene is expressed in the post-mitotic myotubes it is lethal, whereas it is not lethal if expressed in replicating percursor myogenic cells. The viral gene expression at permissive temperature blocks further myogenesis depending on the position of the cells in the myogenic program. The virus does not cancel the replicating, transformed myogenic cells' commitment to, or position in, the myogenic lineage. When the transforming action of the virus is suppressed, the normal myogenic program resumes.
Antibody to terminal deoxynucleotidyl transferase.
Authors: Bollum F J
Published: May 2006

Antibody to homogeneous terminal deoxynucleotidyl transferase (EC 2.7.7.31, nucleoside triphosphate: DNA deoxynucleotidylexotransferase) from calf thymus gland has been prepared using crosslinked antigen derivatives in rabbits. The antisera produced precipitate and neutralize homologous antigen, and crossreact with human, chicken, mouse, and rat terminal transferase. Antiserum to pure terminal transferase provides greatly expanded analytical capabilities for further studies on terminal transferase biology and structure.
Purification of RNA-instructed DNA polymerase from human leukemic spleens.
Authors: Witkin S S , Ohno T , Spiegelman S
Published: May 2006

Particles possessing a density of 1.16 g/ml and encapsulating a 70S RNA and a RNA-instructed DNA polymerase (reverse transcriptase) have been prepared from the spleen of a patient with chronic lymphocytic leukemia. These particles have been converted to cores with a density of 1.26 g/ml and containing the enzyme-RNA complex, in complete analogy to the known RNA tumor viruses of avian and murine origin. The reverse transcriptase was purified from the cores by column chromatography to a stage showing a single major protein band of 70,000 daltons in a gel electrophoresis. The enzyme was capable of transcribing heteropolymeric RNA into DNA complements as demonstrated by specific back hybridization to template RNA. The leukemic spleen would appear to represent an important source of this enzyme, as well as other potentially important leukemia-specific reagents.
Anatomy of herpes simplex virus DNA: evidence for four populations of molecules that differ in the relative orientations of their long and short components.
Authors: Hayward G S , Jacob R J , Wadsworth S C , Roizman B
Published: May 2006

Intact DNA molecules extracted from HSV-1 (herpes simplex virus 1, human herpes virus 1) strain MP virions have a molecular weight of approximately 97 X 10(6), but cleavage with the HinIII restriction enzyme yields fourteen fragments with summed molecular weights of 160 X 10(6). Six "major" fragments occur once in every molecule in the population and account for 60% of the genetic information. Four "minor" fragments are present in amounts equivalent to one copy for every two genomes (0.5 molar ratio) and the other four occur only once in every four molecules (0.25 molar ratio). The minor fragments can be arranged into four equimolar sets, each with summed molecular weights that account for the remaining 40% of the genome. Treatment with lambda 5' exonuclease revealed that all molecules contain 0.5 molar ratio fragments at both termini. These observations and the results of similar analyses of the EcoRI and double HinIII/EcoRI digests indicate that there are four distinct structural forms of HSV DNA which differ only in the relative orientations of two subregions, designated L and S. The L and S segments consist of 82 and 18% of the sequences, respectively, and each has inverted terminally redundant regions that correspond to the internal duplications observed by electron microscopy. The DNA from other strains of HSV-1 and 2 also consists of equal proportions of all four possible permutations of the L and S segments. These unusual features of HSV DNA molecules have novel implications with regard to the genetic map and the mode of replication and evolution of herpes simples viruses.
Allosteric mechanism for codon-dependent tRNA selection on ribosomes.
Authors: Kurland C G , Rigler R , Ehrenberg M , Blomberg C
Published: May 2006

We suggest that the interaction between a codon and its cognate tRNA induces conformational changes in the tRNA. We further suggest that sites on the ribosome preferentially bind tRNA in those conformations which require proper matching of codon and anticodon. According to this model, the codon functions as an allosteric effector which influences the conformation at various sites in the tRNA. This is made possible by the ribosome, which we suggest traps tRNA molecules in those conformation states that maximize the energy difference between cognate and noncognate codon-anticodon interactions. Studies of the interactions between tRNA molecules and their cognate codons in the absence of the ribosome have suggested that triplet-triplet interaction between codon and anticodon is far too weak to account for the specificity of the tRNA selection mechanism during protein synthesis. In contrast, we suggest that such affinity measurements do not adequately describe the interaction between a codon and its cognate tRNA. Thus, such experiments can not detect conformational changes in the tRNA, and, in particular, those stabilized by the ribosome.
Characterization of DNA condensates induced by poly(ethylene oxide) and polylysine.
Authors: Laemmli U K
Published: May 2006

High-molecular-weight DNA is known to collapse into very compact particles in a salt solution containing polymers like poly(ethylene oxide) [(EO)n] or polyacrylate. The biological relevance of this phenomenon is suggested by our recent finding that high concentrations of the highly acidic internal peptides found in the mature T4 bacteriophage head, as well as poly(glutamic acid) and poly(aspartic acid), can collapse DNA in a similar manner. The structure of DNAs collapsed by various methods has been studied with electron microscope. We find (EO)n collapses T4 or T7 bacteriophage DNA into compact particles only slightly larger than the size of the T4 and T7 head, respectively. In contrast, polylysine collapses DNA into different types of structures. Double-stranded DNA collapsed with (EO)n is cut by the single-strand specific Neurospora crassa endonuclease (EC 3.1.4.21) into small fragments. Extensive digestion only occurs above the critical concentration of polymer required for DNA collapse, demonstrating the (EO)n-collapsed DNA contains enzyme-vulnerable regions (probably at each fold), which are preferentially attacked. The size of the DNA fragments produced by limit-digestion with the nuclease ranges between 200 and 400 base pairs when DNA is collapsed by (EO)n. Only fragments of DNA which are larger than 600 base pairs are cut by the endonuclease in (EO)n-containing solution.
Quaternary constraint in hybrid of aspartate transcarbamylase containing wild-type and mutant catalytic subunits.
Authors: Gibbons I , Flatgaard J E , Schachman H K
Published: May 2006

Unusual quaternary constraint in the regulatory enzyme, aspartate transcarbamylase (aspartate carbamoyltransferase or carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2) from Escherichia coli, was demonstrated with a hybrid composed of one inactive "catalytic" subunit from a mutant strain and one active catalytic subunit and three regulatory subunits from the wild-type strain. The hybrid had a high affinity for three molecules of the bi-substrat analog, N-(phosphonacetyl)-L-aspartate, compared to the six strong binding sites in the wild-type enzyme and none in the mutant. However, the Vmax of the hybrid was only about 25% that of the wild-type enzyme. In addition, the hybrid exhibited a very low apparent affinity for the substrate, aspartate [Michaelis constant (Km) about 90 mM], as compared to the wild-type enzyme (apparent Km of 7 mM). No homotropic effect was observed for the hybrid in the absence of nucleotides as contrasted to the cooperativity of the wild-type enzyme; also, large changes in the Vmax of the hybrid were caused by the addition of the nucleotide effectors, CTP and ATP, which do not affect the Vmax of the wild-type, but influence only the cooperativity and the apparent Km. Although the hybrid undergoes a ligand-promoted conformational change analogous to that of the wild-type enzyme, this transition required a 20-fold higher concentration of the substrate analog, succinate. It appears that the "paralysis" of the wild-type catalytic subunit in the hybrid can be attributed to subunit interactions which constrain the molecule in a low-affinity state.
Guanosine 5'-diphosphate 3'-diphosphate (ppGpp): positive effector for histidine operon transcription and general signal for amino-acid deficiency.
Authors: Stephens J C , Artz S W , Ames B N
Published: May 2006

Maximal expression of the histidine operon of Salmonella typhimurium in a coupled in vitro transcription-translation system is strongly dependent upon addition of guanosine 5'-diphosphate 3'-diphosphate (ppGpp). This requirement for ppGpp is exerted at the level of transcription through a mechanism distinct from the his-operon-specific regulatory mechanism. In vivo derepression of the his operon is markedly defective when histidine starvation is imposed on a relA mutant--unable to rapidly increase synthesis of ppGpp--growing in amino-acid-rich medium. Increased sensitivity of relA mutants to growth inhibition by a number of amino-acid analogs suggests that ppGpp is generally important in adjusting expression of amino-acid-producing systems. Analysis of these findings leads us to propose that ppGpp is a positive effector in a system that enables the cell to balance endogenous amino-acid production with environmental conditions of amino-acid availability, and to compensate efficiently for transient changes in these conditions. We propose a unifying theory of the role of ppGpp as the general signal molecule (alarmone) in a "super-control" which senses an amino-acid deficiency and redirects the cell's economy in response.
Fusion of dipalmitoylphosphatidylcholine vesicle membranes induced by concanavalin A.
Authors: van der Bosch J , McConnell M
Published: May 2006

The temperature dependence of fatty acid spin label resonance spectra and freeze fracture micrographs of sonicated dipalmitoylphosphatidylcholine vesicles in the absence and presence of concanavalin A demonstrate a strong interaction of concanavalin A with these lipid membranes, which results in fusion of the vesicles. The rate of this reaction as followed with use of magnetic resonance exhibits a pronounced maximum at 36 degrees, the midpoint of the phase transition range of dipalmitoylphosphatidylcholine vesicles. This maximum is discussed in terms of structural fluctuations, which are maximal in the phase transition range of the membranes.
Contraction of isolated smooth muscle cells by inophore A23187.
Authors: Murray J J , Reed P W , Fay F S
Published: May 2006

Ionophore A23187 causes contraction of smooth muscle cells isolated from the stomach muscularis of Bufo marinus. The peak response is reached within 10 sec of inophore addition and then decays to a lesser, sustained contraction. The initial phase of contraction is maximal at A23187 concentrations greater than 5 X 10(-6) M and half maximal at 5 X 10(-7) M. Chelation of extracellular calcium by ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA) does not affect the initial contraction produced by A23187, whereas it inhibits completely the sustained, secondary phase. Addition of calcium in excess of EGTA restores this phase rapidly and fully to control levels of contraction. A23187 produces a maximal, rapid contraction of muscle cells previously depolarized by incubation in high potassium medium. The ionophore also contracts isolated strips of stomach smooth muscle. A23187 appears to produce contraction of the isolated smooth muscle cells by transmembrane transport of calcium into the cytoplasm from both intracellular and extracellular sources.
Turning on of activities in unfertilized sea urchin eggs: correlation with changes of the surface.
Authors: Mazia D , Schatten G , Steinhardt R
Published: May 2006

Unfertilized sea urchin eggs exposed to low concentrations of ammonia enter into a number of activities which normally appear after fertilization. It is shown that the effects are attributable to ammonia, rather than to NH4+ ions of elevated pH. The same effects are obtained by exposure to isotonic urea and to glycerol at very low ionic strengths. All treatments which produce these changes (such as the turning on of chromosome replication and condensation in unfertilized eggs) also bring about changes of the outer cell surface which are visible in the scanning electron microscope. The most striking indicator is the elongation of the microvilli which cover the surface of the unfertilized egg. The changes of the surface are interpreted as the dissociation of a component from the outer surface layer. This component is not the "vitelline" sheet as defined morphologically or by the ability of the egg to form a fertilization membrane upon insemination. It is proposed further that this component is a peripheral component of the plasma membrane, whose removal modifies the membrane functionally and leads to the derepression of various processes within the egg.
Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells.
Authors: Wang K , Ash J F , Singer S J
Published: May 2006

A new high-molecular-weight protein, named filamin, was isolated from chicken gizzard. In chicken gizzard, filamin is present in an amount approximately 30-40% of that of myosin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of highly purified filamin revealed a single polypeptide of about 250,000 daltons. Rabbit antibody directed against purified chicken gizzard filamin did not crossreact with myosin purified from the same source. By the use of microcomplement fixation and indirect immunofluorescent staining with antibodies to chicken gizzard filamin, an antigenically similar or identical protein was found to be widely distributed both in other organs of the chicken and in cultured cells of other species, but not in chicken skeletal muscle. In cultured cells, filamin was found largely to be arranged as a filamentous array very similar to that found for myosin. These data imply that filamin is a widely occurring and chemically conserved component of filaments is smooth muscle and non-muscle cells.
Hydrogen bonding in yeast phenylalanine transfer RNA.
Authors: Quigley G J , Wang A H , Seeman N C , Suddath F L , Rich A , Sussman J L , Kim S H
Published: May 2006

Further analysis of the three-dimensional electron density map of yeast phenylalanine tRNA is presented. Attention is focused on the several types of unique hydrogen bonding that are found in the molecule and a number of sections of the electron density map are presented. These sections are compared with an electron density map of a dinucleoside phosphate. The bases in the helical stem regions are all involved in Watson-Crick hydrogen bonding interactions with the exception of the guanine-uracil base pair. Several additional tertiary hydrogen bonding interactions are described.
Primary charge separation in bacterial photosynthesis: oxidized chlorophylls and reduced pheophytin.
Authors: Fajer J , Brune D C , Davis M S , Forman A , Spaulding L D
Published: May 2006

Bacteriopheophytin, the magnesium-free base of bacteriochlorophyll, undergoes reversible one-electron reduction in organic solvents to yield an anionic free radical with characteristic optical and electron spin resonance spectra. The reduction potential of bacteriopheophytin, E1/2 approximately --0.55 V against a normal hydrogen electrode, compared to E1/2 approximately --0.85 V for bacteriochlorophyll, renders it a likely electron acceptor in the primary charge separation of photosynthesis. Comparison of these data with picosecond optical changes recently observed upon pulsed laser excitation of bacterial reaction centers leads us to propose that bacteriopheophytin is indeed a transient electron acceptor and that the primary charge separation of bacterial photosynthesis occurs between the bacteriochlorophyll complex P870 and bacteriopheophytin to yield the radicals of the oxidized chlorophyll dimer cation and reduced pheophytin anion.
Regulation of ferredoxin-catalyzed photosynthetic phosphorylations.
Authors: Arnon D I , Chain R K
Published: May 2006

Under aerobic conditions that are likely to prevail in chloroplasts in vivo, the optimal concentration of ferredoxin for cyclic photophosphorylation was found to be equal to that required for NADP reduction and about one-tenth of that needed for cyclic photophosphorylation under anaerobic conditions. In the presence of ferredoxin and NADP, cyclic photophosphorylation operated concurrently with noncyclic photophosphorylation, producing an ATP: NADPH ratio of about 1.5. The effective operation of ferredoxin-catalyzed cyclic photophosphorylation by itself required a curtailment of the electron flow from water which was accomplished experimentally by the use of either an inhibitor or far-red monochromatic light. An unexpected discovery was that the operation of cyclic photophosphorylation by itself was also regulated by a back reaction of NADPH and ferredoxin with two components of chloroplast membranes, component C550 and cytochrome b559. The significance of these findings to photosynthesis in vivo is discussed.
Differences among myosins synthesized in non-myogenic cells, presumptive myoblasts, and myoblasts.
Authors: Chi J C , Fellini S A , Holtzer H
Published: May 2006

Myosins synthesized in non-myogenic cells and replicating presumptive myoblasts differ from those synthesized in postmitotic mononucleated myoblasts and myotubes. Myoblasts and myotubes synthesize the definitive light chains, MLC1 and MLC2. These light chains display different molecular weights in sodium dodecyl sulfate-polyacrylamide gels from the fibroblast light chains FLC1 and FLC2 synthesized in non-myogenic cells and presumptive myoblasts. There are immunological differences between the myosin heavy chains synthesized in myoblasts and myotubes and those synthesized in non-myogenic cells and presumptive myoblasts. Fluorescein-labeled antibodies against skeletal light meromyosin are bound only along the lateral edges of emerging and definitive A-bands. This antibody to light meromyosin is not bound to the outside of, or the microfilaments subtending, the plasma membrane in non-myogenic cells or in myoblasts or in myotubes. These findings suggest that: (1) non-myogenic cells and replicating presumptive myoblasts synthesize similar myosin heavy and light chains; (2) replicating presumptive myoblasts synthesize a different set of myosins from those synthesized by their postmitotic daughters, the myoblasts; (3) the myosins associated with the plasma membranes of non-myogenic and myogenic cells are products of structural genes distinct from those coding for the myosins for skeletal myofibrils.
Snythesis and differentiation of plasma proteins in cultured embryonic chicken liver cells: a system for study of regulation of protein synthesis.
Authors: Grieninger G , Granick S
Published: May 2006

A new system is described for studying the control of protein synthesis. In a monolayer culture of chick embryo liver cells, plasma proteins are synthesized for three days at in vivo rates. The plasma proteins are secreted into the culture medium and without concentration are detected there simply and sensitively by a modified Laurell electronimmunoassay. Secretion of the newly synthesized plasma proteins occurs within 30 min of their synthesis. Thus, rates of synthesis of the plasma proteins can be followed readily from rates of their accumulation in the culture medium. This system has the following advantages for the study of protein synthesis: cells do not have to be disrupted for the assay; the cell population can be followed over several days; it is not necessary to label the proteins radioactively; and turnover of plasma proteins is negligible and need not be taken into account. The usefulness of the system is illustrated by a number of findings. The spectrum of plasma proteins synthesized in culture changed qualitatively and quantitatively. Albumin synthesis steadily decreased with culture time and stopped at the third day, whereas the synthesis of some new plasma proteins ("adult") was induced. These qualitative changes suggest differential gene expression in culture and a special control of albumin synthesis in vivo, different from the synthesis of the other plasma proteins. Quantitative changes in the rates of synthesis of specific plasma proteins suggest a competition among their messenger RNAs for components of the translational machinery. Insulin has a differential effect on the synthesis of specific plasma proteins at concentrations within the physiological range of the hormone.
RNA-primed DNA synthesis: specific catalysis by HeLa cell DNA polymerase alpha.
Authors: Spadari S , Weissbach A
Published: May 2006

We have analyzed and compared the responses of the three major HeLa cell DNA polymerases (alpha, beta, and gamma) to a HeLa DNA template with short RNA or DNA primers hybridized to it. Only DNA polymerase alpha is able to synthesize DNA covalently bonded to the RNA primer via a 3' yields 5' phosphodiester bond. 32P transfer experiments showed that all combinations of ribo- and deoxyribonucleotides are represented in the RNA-DNA linkages but their distribution is nonrandom. The RNA-DNA linked molecules base-paired to a HeLa DNA template strand represent a possible "natural" in vitro primer-template for DNA polymerases and can be extended by all three DNA polymerases (alpha, beta, and gamma). These findings indicate that DNA polymerases beta and gamma are capable of DNA-primed but not RNA-PRIMED DNA synthesis, while DNA polymerase alpha is capable of both RNA-primed and DAN-primed DNA synthesis.
Effects of cytochaslasin B and colcemide on myogenic cultures.
Authors: Holtzer H , Croop J , Dienstman S , Ishikawa H , Somlyo A P
Published: May 2006

Muscle cultures treated with cytochalasin B yield mono- and oligonucleated cells of two kinds: (i) arborized, replicating precursor myogenic cells and fibroblasts; and (ii) round, post-mitotic, terminally differentiating myoblasts and myotubes. The arborized cells do not bind fluorescein-labeled antibody against myosin, do not contract rhythmically, and do not display hexagonally stacked thick and thin filaments. The round, mono-nucleated myoblasts and round, oligonucleated myotubes bind the fluorescein-labeled antibody against myosin, contract rhythmically, and display clusters of hexagonally-stacked thick and thin filaments. When cytochalasin B is removed and replaced by colcemide, the arborized cells, but not the post-mitotic muscle cells, acquire a radial symmetry and are induced to assemble massive, meandering cables that may occupy over 25% of the cell volume. These tortuous calbes are positively birefringent and consist exclusively of enormous numbers of 100-A, intermediate-sized filaments.
Regulation of development in Myxococcus xanthus: effect of 3':5'-cyclic AMP, ADP, and nutrition.
Authors: Campos J M , Zusman D R
Published: May 2006

An assay was developed to study the regulation of fruiting in Myxococcus xanthus. The nucleotides, adenosine 3':5'-cyclic monophosphate (cyclic AMP) and adenosine diphosphate (ADP), were found to greatly stimulate fruiting under the assay conditions. Very sharp concentration optima were observed. Even under conditions of starvation, these nucleotides greatly increased the number of aggregation sites. Nutrition was found to influence fruiting body morphology. The effect of amino acids on the nucleotide stimulation of fruiting was studied under our assay conditions. L-Methionine and L-isoleucine (1 mM) completely blocked either L-threonine or D,L-diaminopimelic acid synergistically enhanced the amount of fruiting in the presence of these nucleotides. The data presented suggest the existence of differentiation-related regulatory compounds in M. xanthus.
Properties of Molten Magnesium Oxide
Authors: Leu An-Lu , Ma Shao-Mu , Eyring Henry
Published: May 2006

The Significant Structure Theory of Liquids has been used to calculate the thermodynamic properties, viscosity, self-diffusion coefficient, and specific conductance of molten magnesium oxide, taking account of the decomposition of MgO to Mg and O
2
species in the gas-like part of the partition function.
Cyclic AMP and cyclic GMP concentrations in serum- and density-restricted fibroblast cultures.
Authors: Moens W , Vokaer A , Kram R
Published: May 2006

Mouse fibroblasts transformed by simian virus 40 (SV3T3 cells) are characterized by cyclic AMP and cyclic GMP levels, respectively, about half and twice those found in growing untransformed 3T3 cells. Density-dependent inhibition of growth is correlated with reduced cyclic GMP concentrations in 3T3 and four different density-restricted revertant lines derived from SV3T3. The levels of cyclic AMP are not increased at confluence. Upon serum restriction, serum-dependent cell lines show a greater increase in intracellular cAMP than serum-insensitive lines. Cyclic GMP levels are greatly reduced, even in serum-insensitive density revertants, but not in SV3T3. Serum readdition to all serum-dependent lines is followed by a rapid decrease in cyclic AMP and increase in cyclic GMP concentrations. The magnitude of these responses is decreased in SV3T3 and density revertants.