Antithrombin III binds to human platelets

The receptor site for antithrombin III (AT III) was investigated in normal human platelets. [¹²⁵I] iodinated AT III was utilized as tracer for the binding assay. Equilibrium of AT III binding was reached within 2 min. The binding capacity was pH-dependent with the optimum around pH 7.0. Binding specificity was demonstrated by inhibition of [¹²⁵I] AT III ligation using an excess amount of non-labeled AT III. The AT III·heparin complex did not supress [¹²⁵I] AT III binding. Analysis of binding data by Scatchard plot revealed a single class of binding sites with K_d of 3.2 × 10^?7 M and binding capacity of 3840 per platelet.     

GABA induces behavioral and developmental metamorphosis in planktonic molluscan larvae

Swimming planktonic larvae of the marine gastropod mollusc Haliotis rufescens require exogenous GABA or its homologs for induction of their genetically programed behavioral and developmental metamorphosis to the adult form. This requirement is stereochemically specific and absolute; GABA at 10(-6) M is fully effective in the induction of cellular differentiation, proliferation and organogenesis. The kinetics of the development of larval competence for GABA induction, and of the early metamorphic processes induced by GABA, are described. Biochemical, histological and electron micrographic analyses suggest that cyclic AMP, calcium, and a glycopeptide secretion from the cephalic sensory complex may mediate transduction of the GABA signal in the control of behavioral and morphogenetic changes induced by this environmentally deployed transmitter substance. This first observation and characterization of a major role for GABA in the control of differentiation and development, and the experimentally tractable system in which these are demonstrated, are of significance for further biomedical research.     

Triphasic concentration effects of gentamicin on activity and misreading in protein synthesis.

Gentamicin is shown to exert a triphasic concentration effect on peptide synthesis in vitro with natural messengers. Low concentrations (up to 2 micron) caused slowing and a decrease in total synthesis, but little misreading (assayed with extracts lacking Glu-tRNA); the inhibition was greater with an initiating system (with phage RNA as messenger) than with pure chain elongation on purified endogenous polysomes of Escherichia coli. Moderate concentrations (up to 100 micron) slowed synthesis less, markedly increased its duration in the noninitiating system, and strongly stimulated misreading; at optimal concentrations total synthesis was even greater than normal. Moreover, with phage RNA these concentrations increased the synthesis of large polypeptides. We conclude that binding of gentamicin to its first site causes inhibition but little misreading; binding to additional site(s) partly reverses the inhibition by first-site binding and markedly stimulates misreading, and the misreading appears to favor "readthrough" of termination codons. In the third phase (greater than 100 micron) synthesis is slowed again but the pattern of misreading does not appear to be altered; this effect need not involve a specific further action on the ribosome.     

The effect of growth hormone on the metabolism of a tryptophan load in the liver and brain of hypophysectomized rats.

Growth hormone antagonizes the induction of tryptophan pyrrolase and tyrosine amino-transferase by cortisol. We have shown that contrary to previous reports, growth hormone is also capable of antagonizing the induction of these enzymes by tryptophan and alpha-methyl tryptophan. As alpha-methyl tryptophan is not metabolized appreciably in the rat, our data show that growth hormone does not act indirectly through changes in the liver tryptophan content as was suggested previously. Growth hormone decreases the rate of tryptophan catabolism in vivo after induction of tryptophan pyrrolase by tryptophan and alpha-methyl tryptophan. Because the rate of catabolism of a tryptophan is slower in animals treated with growth hormone, tissue tryptophan levels and the rate of synthesis of 5-hydroxyltryptamine in the brain are higher in these animals than in those receiving tryptophan alone. Thus, although tryptophan administration raises brain 5-hydroxytryptamine levels, induction of tryptophan pyrrolase in the liver, by the load, limits the extent and duration of the rise in brain 5-hydroxytryptamine synthesis. This has important implications for the clinical use of tryptophan in psychiatric disorders, where tryptophan is given to produce long-lasting elevations of brain 5-hydroxytryptamine.     

Enzyme Polymorphism and Cyclic Parthenogenesis in DAPHNIA MAGNA. I. Selection and Clonal Diversity

Genotype frequencies and fecundities were recorded over a period of two years for three polymorphic enzyme loci (Est, Mdh and Got) in a parthenogenetic natural population of Daphnia magna Straus (Crustacea: Cladocera). There was a large excess of heterozygotes at each locus, and some nonrandom association between loci, although 29 different three-locus genotypes were detected. There were small but significant changes in genotype frequencies that did not follow any clear seasonal cycles or overall trends, and the genotypes often differed significantly in fecundity, although the direction of the difference was not constant. These fitness differences were probably not attributable to the specific loci studied.--Models of balancing selection are of two types: segregation-balanced (e.g., heterosis) and competition-balanced (e.g., frequency dependence). Only the latter type can stabilize diversity in a clonal population. The observed selection was not heterotic, but it is not certain that it was stabilizing either. Clonal competition did not lead to victory by a single, fittest clone; genotypic diversity remained high.     

Isolation and partial characterization of a new soluble b-type cytochrome (b9) from rat liver.

A new soluble cytochrome, designated as cytochrome b9, was purified to apparent homogeneity from rat liver. The absorption maximum of the oxidized (the native form) cytochrome b9 at room temperature was 413 nm. The dithionite-reduced cytochrome b9 had absorption maxima at 556, 527, and 423 nm. The prosthetic group of cytochrome b9 was identified as protoheme IX. From gel filtration experiments, the molecular weight of cytochrome b9 was estimated to be 125,000. Polyacrylamide gel electrophoresis experiments in the presence of sodium dodecyl sulfate showed that the molecular weight of its subunit was 61,000. The native form of cytochrome b9 was thus a dimer. The amount of heme/mol of dimer was 3.3 mol. Cytochrome b9 was autoxidizable and did not bind CO, 2.2 mM cyanide, or 2.2 mM azide. On the basis of its molecular weight of 125,000, the millimolar extinction coefficients of dimeric cytochrome b9 at 280 and 413 nm were 384 and 380, respectively. The absorbance at 280 nm/mg cytochrome b9 was 3.1. Cytochromes b9 and H-450 (I.-C. Kim and W.C. Deal (1976) Biochemistry 15, 4925-4930) are the only b-type, soluble cytochromes which have been isolated from mammalian liver; they are not found in tissues of heart, lung, kidney, and brain. The biological function of cytochrome b9 was not determined.