Role of methyl groups in dynamics and evolution of biomolecules

Recent studies have discovered strong differences between the dynamics of nucleic acids (RNA and DNA) and proteins, especially at low hydration and low temperatures. This difference is caused primarily by dynamics of methyl groups that are abundant in proteins, but are absent or very rare in RNA and DNA. In this paper, we present a hypothesis regarding the role of methyl groups as intrinsic plasticizers in proteins and their evolutionary selection to facilitate protein dynamics and activity. We demonstrate the profound effect methyl groups have on protein dynamics relative to nucleic acid dynamics, and note the apparent correlation of methyl group content in protein classes and their need for molecular flexibility. Moreover, we note the fastest methyl groups of some enzymes appear around dynamical centers such as hinges or active sites. Methyl groups are also of tremendous importance from a hydrophobicity/folding/entropy perspective. These significant roles, however, complement our hypothesis rather than preclude the recognition of methyl groups in the dynamics and evolution of biomolecules.     

Amine oxidase in unicellular microorganismsMethanosarcina barkeri andTetrahymena pyriformis

A comparison has been performed of catalytic properties of unicellular microorganism amine oxidases (AO) from two new enzyme sources, the bacteriumMethanosarcina barkeri and the infusoriaTetrahymena pyriformis. It was shown that the both studied AO deaminate tyramine, serotonin, and benzylamine, but do not deaminate histamine. The AO fromMethanosarcina barkeri catalyzes deamination of all three substrates at an identical rate, while the rate of tyramine deamination under effect of AO fromTetrahymena pyriformis is one order higher than the rate of serotonin deamination, and about two orders higher than the rate of benzylamine deamination. Based on the data of the substrate-inhibitor analysis, a suggestion was made about the existence of one center for the substrate binding in the AO of the studied bacterium, while several centers in the AO of the studied infusoria.     

Modeling Tetrahymena thermophila growth and protease production

Oxygen concentrations stimulated growth (maximum number of cells) and protease secretion by Tetrahymena thermophila. Agitation and aeration conditions for growth and protease secretion were optimised by a central composite design. The best optimised combination was a stirrer speed of 338 rpm and an aeration of 1 vvm. Journal of Industrial Microbiology & Biotechnology (2000) 25, 58–61. Received 24 September 1999/ Accepted in revised form 06 March 2000     

Red-breasted goose colonies on the Taimyr Peninsula: Factors responsible for the proximity of goose nests to nests of peregrine falcons,rough-legged buzzards,and snowy owls

Red-breasted goose colonies have been studied near Medusa Bay (73°21′N, 80°32′E), on the northwestern Taimyr Peninsula, and along the Agapa River (70°11′N, 86°15′E) down to its mouth (71°26′N, 89° 13′E), in the central Taimyr Peninsula. Red-breasted geese nesting near peregrine falcons are protected by the falcons from arctic foxes; however, they are sometimes attacked by the falcons themselves. In the colonies near peregrine falcon nests, the vast majority of goose nests were situated no farther than 100 m from the falcon nest. When food is abundant, falcons protect a larger area around their nest. The distance between the falcon nest and the surrounding goose nests is inversely related to the falcon’s activity. In years of higher falcon activity, falcons prevent red-breasted geese from nesting as close to their nest as in years of lower falcon activity. Additional stimuli are required for red-breasted geese to form colonies near rough-legged buzzard nests. The distance between snowy owl nests and red-breasted goose nests was smaller when arctic foxes were abundant than when they were scarce.     

Characterization of a transferrin-diphtheria toxin conjugate

We report here the synthesis and properties of a hybrid toxin prepared by covalently coupling diphtheria toxin to transferrin. The purified material contained two major hybrid protein species and was highly cytotoxic to mouse LMTK- cells in culture, reducing protein synthesis by 50% in 24 h at a concentration of 1 ng/ml. Cytotoxic activity was completely abolished in the presence of exogenous transferrin or anti-transferrin or anti-diphtheria toxin, thus demonstrating that the hybrid toxin was intoxicating cells via their transferrin receptors and that both the diphtheria toxin and transferrin components of the conjugate were necessary for activity. NH4Cl, a drug that elevates the pH within acidic intracellular vesicles, also blocked cytotoxic activity, suggesting that a low intravesicular pH was required for activity. The inhibitory effect of NH4Cl could be abolished by exposing toxin-treated cells to acidic culture medium, further implicating an acid-dependent step in the mechanism of the hybrid toxin action. Studies on the kinetics of intoxication also implied that endocytosis and exposure to a low pH within vesicles were necessary for cytotoxicity. Altogether, the results suggest that the transferrin-diphtheria toxin conjugate binds to transferrin receptors and is internalized into acidic endocytic vesicles. The enzymatic moiety of diphtheria toxin then apparently enters the cytosol in response to the low pH and subsequently arrests protein synthesis.     

Immunocytochemical detection of triphosphoinositide in vestibular hair cells

Triphosphoinositide (TPI), an aminoglycoside receptor and a possible regulator of cationic permeation through its ability to bind with Ca++, was localized by the protein-A gold technique in vestibular sensory epithelia using an antibody highly specific to TPI. TPI was detected on the stereocilia, kinocilia, and cuticular plate of hair cells, and in the reticular membrane of supporting cells. The cilia of hair cells are damaged by aminoglycosides at a relatively early stage of toxicity. Ca++-regulated bioactivity in this area is probably involved.     

Synthesis of new olean-18(19)-ene derivatives from allobetulin

New olean-18(19)-ene triterpenoids were effectively synthesized by the interaction of allobetulin or its acetate with phosphorous oxychloride in refluxing pyridine. The structures of the synthesized 17-chloromethyloleane-18(19)-enes were confirmed by NMR spectroscopy and X-ray analysis.