Tubulin subunit carboxyl termini determine polymerization efficiency

Cleavage of tubulin by subtilisin removes a small (Mr less than 2000) fragment from the C-terminal end of both alpha and beta subunits. The resulting protein is much reduced in negative charge. The cleaved, less acidic protein retains its competence to polymerize in a GTP-dependent and cold-, GDP-, and podophyllotoxin-sensitive manner and assembles into sheets or bundles of twisted filaments. The critical concentration for polymerization of the cleaved protein is about 50-fold lower than that for intact tubulin. It is proposed that the C termini of the subunits normally impede polymerization.     

Emergence of blackflies (Diptera: Simuliidae) from a small forested stream in Ontario

Three species of blackflies were found in emergence-trap samples taken overa period of 8 months from a second-order, forested, cold-stenothermal stream in southern Ontario. The emergence phenologies of the two common species, Prosimulium mixtum and Stegopterna mutata, are described and compared with their phenologies in other streams in North America. Hypotheses are presented for the poor faunal diversity and prolonged emergence of the two species in the stream studied. Emergence data are used to describe the pupal distribution in the stream.Wing-length measurements showed a distinct dimorphism in P. mixtum: females were larger than males. Adult size (except of P. mixtum females) varied among sampling sites in the stream and, in the case of S. mutata, this variation was time dependent.     

Structural localization of the sequence alpha 235-242 of the nicotinic acetylcholine receptor

Two monoclonal antibodies (mAb 254 and 255) were obtained against a synthetic peptide corresponding to the sequence 235-242 of the alpha-subunit of Torpedo acetylcholine receptor. These mAbs could bind to receptor in native membrane vesicles only when these vesicles were permeabilized, suggesting that the sequence alpha 235-242 is exposed on the cytoplasmic surface of the receptor. Further evidence for the cytoplasmic localization of this sequence was partial competition for binding between these mAbs and mAbs previously demonstrated to bind to the cytoplasmic part of the receptor. A model is proposed which accounts for all the experimental data obtained thus far on the transmembrane orientation of the subunit polypeptide chains.     

The role of histones H3 and H1 in the formation of thymine-lysine cross-links during UV-irradiation of deoxyribonucleoprotein

The effect of UV light (lambda = 254 nm) on calf thymus DNP at low ionic strengths was studied. It was found that at the irradiation doses used the protein in the DNA-protein complex increases as the irradiation dose rises. Thermal treatment and acid hydrolysis resulted in a predominant release of histones H3 and H1 from the complex. Data from liquid high performance chromatography, amino acid analysis, thin-layer chromatography point to the induction by UV-light of a thymine-lysine bond, whose formation involves DNA thymines and histone lysine residues, predominantly H3 and H1 fractions.     

Effect of nucleoside triphosphates on cyclic nucleotide phosphodiesterase: role of protein modulators

The effects of nucleoside triphosphates (ATP and GTP) on phosphodiesterase (PDE) of brain and outer segments of the retina enriched or devoid of protein modulators were studied. In the case of retinal outer segment PDE the enzyme activity was considerably inhibited by both nucleosides only when the enzyme was separated from the inhibitor. In case of brain PDE, on the contrary, the effect of the nucleosides was much more pronounced in the enzyme preparation coupled with the protein activator, calmodulin. The latter when added to brain PDE devoid of the activator in the presence of ATP and GTP considerably reduced the enzyme activity. An addition of the inhibitor simultaneously with GTP to the purified PDE of outer segments increased the PDE activity. The constants for the inhibition of brain PDE coupled with calmodulin and retinal outer segment PDE separated from the inhibitor by ATP and GTP were determined.     

Susceptibility of human and murine Chlamydia trachomatis serovars to granulocyte- and epithelium-derived antimicrobial peptides.

Four antimicrobial peptides, protegrin-1, RTD-1, cryptdin-4, and indolicidin, were tested for their ability to inhibit the in vitro growth of Chlamydia trachomatis serovars E, L2, and mouse pneumonitis (MoPn). In general, protegrin-1 was found to have the strongest anti-chlamydial activity. Overall, of the three serovars tested, L2 was the most susceptible while MoPn was the most resistant to these peptides.     

Activity of cAMP-dependent protein kinases and cAMP-binding proteins from rat renal cytosol upon dehydration

The activity of cAMP-dependent protein kinases, cAMP binding and the spectrum of cAMP-binding proteins in renal papillary cytosol of intact rats and of rats kept on a water-deprived diet for 24 hours were investigated. It was found that the stimulation of protein kinases by 10(-6) M cAMP in the experimental group was significantly higher than in the control one. On DEAE-cellulose chromatography, the position of peaks of the specific cAMP binding corresponded to those of the regulatory cAMP-dependent protein kinases type I and II. Under these conditions, more than 80% of the binding activity in intact animals was localized in peak II, whereas in rats kept on a water-deprived diet over 60% of the binding activity was localized in peak I. The total binding activity of cytosol in experimental animals remained unchanged is compared to intact rats. It is suggested that in renal papilla dehydration is accompanied by the induction of synthesis of regulatory subunits of cAMP-dependent protein kinase type I.