The heparin binding domain of S-protein/vitronectin binds to complement components C7, C8, and C9 and perforin from cytolytic T-cells and inhibits their lytic activities

S-Protein/vitronectin is a serum glycoprotein that inhibits the lytic activity of the membrane attack complex of complement, i.e., of the complex including the proteins C5b, C6, C7, C8, and C9n. We show that intact S-protein/vitronectin or its cyanogen bromide generated fragments also inhibit the hemolysis mediated by perforin from cytotoxic T-cells at 45 and 11 microM, respectively. The glycosaminoglycan binding site of S-protein/vitronectin is responsible for the inhibition, since a synthetic peptide corresponding to a part of this highly basic domain (amino acid residues 348-360) inhibits complement- as well as perforin-mediated cytolysis. In the case of C9, the synthetic peptide binds to the acidic residues occurring in its N-terminal cysteine-rich domain (residues 101-111). Antibodies raised against this particular segment react 25-fold better with the polymerized form of C9 as compared with its monomeric form, indicating that this site becomes exposed only upon the hydrophilic-amphiphilic transition of C9. Since the cysteine-rich domain of C9 has been shown to be highly conserved in C6, C7, and C8 as well as in perforin, the inhibition of the lytic activities of these molecules by S-protein/vitronectin or by peptides corresponding to its heparin binding site may be explained by a similar mechanism.     

Changes in fiber composition of soleus muscle during rat hindlimb suspension

Chronic reduction of gravitational load in the rear limbs of rats to simulate the influence of near-zero gravity in skeletal muscles has been shown previously to elicit atrophy in the soleus muscle. Use of this model by the present investigation indicates that soleus atrophy was characterized by a decline in the number of fibers in groups that contained the slow isoenzyme of myosin and which were classified as type I from intensity of staining to myofibrillar actomyosin adenosinetriphosphatase (ATPase) and to NADH tetrazolium reductase. Furthermore total fiber number was not changed, whereas fibers containing the intermediate isoenzyme and those classified as type IIa increased. There results could be explained by either a change in the composition within existing fibers or a simultaneous loss of slow fibers and de novo synthesis of intermediate and fast fibers. Evidence for transformation included an absence of embryonic or neonatal myosin in muscles from suspended rats and the constant fiber number that was unchanged by 4 wk of suspension. Furthermore although fiber areas of both groups of type I and IIa fibers declined during suspension, variability of the fiber areas within each group did not increase.     

Translational discrimination between the four RNAs of alfalfa mosaic virus. A quantitative evaluation

In an attempt to relate the translational characteristics of alfalfa mosaic virus (A1MV) RNAs to their structure [Ravelonandro et al. (1983) Nucleic Acids Res. 11, 2815-2826; Gehrke et al. (1983) Biochemistry 22, 5157-5164] we measured the relative affinities (discrimination ratios) of these RNAs for the initiation complex, in the wheat germ extract and in the nuclease-treated reticulocyte lysate, using a competition method designed by Brendler et al. [(1981) J. Biol. Chem. 256, 11747-11754]. As a prerequisite of this study we ascertained that the molecular mass distribution of the translation products was independent of RNA concentration in both translation systems. In the wheat germ extract the discrimination ratios are very similar for two strains of A1MV (S and B) which differ mainly by the presence (strain S) or absence (strain B) of a stable 5'-proximal hairpin. Hence this structure has no bearing on discrimination. Taking the affinity of RNA 3 as reference, the following orders of magnitude are found for the affinities of the different RNAs in the wheat germ: RNA 3, 1.0; RNA 1, 10; RNA 2, 60; RNA 4, 150. In the reticulocyte lysate the discrimination ratios are not significantly different from the wheat germ. Thus it seems that the mechanism of discrimination is essentially the same in the two translation systems, despite a difference in rate-limitation.     

Cleavage of dT8 and dT8 phosphorothioyl analogues by Escherichia coli DNA topoisomerase I: product and rate analysis

Escherichia coli DNA topoisomerase I catalyzes the cleavage of short, single-stranded oligodeoxynucleotides with dT8 as the shortest cleavable oligo(thymidylic acid). The 5'-32P-labeled products formed from the cleavage of [5'-32P]dT8 are dT5, dT4, and dT3 with over 70% of the substrate cleaved to dT4. Mg(II) ions affect this product distribution by increasing the percentage of dT4 formed. The substitution of a sulfur atom for a nonbridging oxygen atom in a phosphodiester linkage yields oligodeoxynucleotide phosphorothioyl (PS) analogues. The epimers of the analogues were separated, and the position and stereochemistry of the phosphorothiodiester bond were determined. Topoisomerase I is stereospecific in its reactivity toward these analogues. With the oligodeoxynucleotide PS analogue substrates, the rate of cleavage, the stereospecificity, and the product distribution depend upon the position and the stereochemistry of the phosphorothiodiester linkage.     

Glucose and ketone bodies production in hepatocytes isolated from fetuses at term--II. Effect of maternal adrenalectomy

Glucose and ketone bodies production has been studied in hepatocytes isolated from fetuses at term of fed and fasted adrenalectomized mothers. Maternal adrenalectomy diminishes the fetal liver weight. This effect is increased when the adrenalectomized pregnant rat is fasted for the last 2 days of gestation. Maternal adrenalectomy diminishes glucose production in hepatocytes isolated from fetuses at term. This diminution is markedly greater when the adrenalectomized pregnant rat is fasted for the last 48 hr of gestation. Maternal adrenalectomy diminishes ketone bodies production in hepatocytes isolated from fetuses at term.