Relationship between alpha-helical propensity and formation of the ribonuclease-S complex.

Variant semisynthetic ribonuclease-S complexes were characterized in which the helical glutamic acid 9 residue was replaced by either leucine or glycine. The Leu-9 and Gly-9 synthetic peptides, corresponding otherwise to residues 1 through 15 of bovine pancreatic ribonuclease, were studied with respect to the ability to bind, and generate enzymic activity, with the complementary native protein fragment containing residues 21 through 124 of ribonuclease (RNAase-S-(21–124)). Both the Leu and Gly peptides bind to the RNAase-S-(21–124) to yield complexes with catalytic properties similar to those obtained with the Glu-9-containing peptide of residues 1 through 20 of ribonuclease (RNAase-S-(1–20)). However, whereas the binding affinity of Leu peptide to RNAase-S-(21–124) is only a factor of three less than that for RNAase-S-(1–20), that for Gly peptide is about 20-fold less than that for RNAase-S-(1–20). The stronger binding of Leu than Gly peptide corresponds to the observed propensity of leucine but not glycine for the α-helical conformation in globular proteins.In spite of the weakened affinity of the Gly peptide for RNAase-S-(21–124), it is essentially fully as capable as the Leu-9 and RNAase-S-(1–20) peptides in directing the re-formation of correct disulfide-containing conformation of RNAase-S-(21–124) after disulfide randomization of the latter.     

The effect of valinomycin and nigericin on the efficacy of bacteriophage infection of staphylococcal cells

The effect of ionophore antibiotics, valinomycin and nigericin, on the generation of the membrane potential, the pH gradient and the efficacy of phage infection in tetracycline-resistant staphylococci has been studied. Valinomycin at a concentration of 0.5 microM induces the dissipation of the membrane potential, and nigericin at a concentration of 12.0 microM decreases the value of the pH gradient on the membrane of staphylococci. The separate use of antibiotics has no essential influence on the efficacy of phage infection. The combined use of valinomycin and nigerimycin produces the maximum inhibition of phage infection (64.5%) at the stage of the introduction of DNA into the bacterial cell, which is indicative of a definite role played by the membrane potential and the pH gradient in the transport of phage DNA into staphylococcal cells.     

Partial trisomy 11q as the result of sporadic translocation

Summary Partial trisomy 11q due to a sporadic translocation was found in a mentally retarded girl with multiple abnormalities. The proportion of sporadic translocations involved in the total incidence of partial trisomies is discussed.     

Isolation of Tyr- mutants of facultative methylotrophic Pseudomonas sp. M

A method for isolation of Tyr- mutants of facultative methylotrophic bacteria Pseudomonas sp. M which possess two tyrosine synthesis pathways is presented. The method is based on the two-step blocking of the tyrosine synthesis: the first step of the supplementary pathway of synthesis from phenylalanine, the second being the main pathway from 4-hydroxyphenylpyruvate.     

Purification and properties of NADPH-dependent aldehyde reductase from human liver.

An aldehyde reductase (EC 1.1.1.2) from human liver has been purified to homogeneity. The enzyme is NADPH-dependent, prefers aromatic to aliphatic aldehydes as substrates, and is inhibited by barbiturates and hydantoins. The following physicochemical parameters were determined: molecular weight, 36,200; sedimentation coefficient, 2.9 S; Stokes radius, 2.65 nm; isoelectric point, pH 5.3; extinction coefficient at 280 nm, 54,300 M-1 cm-1. Results from polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate, gel filtration, and ultracentrifugation suggest a monomeric structure. On molecule of NADPH binds to the enzyme causing a red shift of the coenzyme absorption maximum from 340 to 352 nm. The amino acid composition has been determined and a partial specific volume of 0.74 was computed from these data. An alpha-helicity of 7 and 18% was estimated from the ellipticities at 208 and 222 nm, respectively. Combination of the most reactive thiol group with p-mercuribenzoate does not cause loss of catalytic activity. Inactivation occurs when more than one thiol group is modified. The presence of NADPH or NADP+ prevents loss of activity by thiol modification. The comparison of structural features of aldehyde reductase with other monomeric and oligomeric dehydrogenases suggest similarities of aldehyde reductase with octopine dehydrogenase.