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Biomedical subjects

M Wilchek

Publications and source records attributed to M Wilchek.

At least 199 records · Page 11Linked to original sources

Reversibility of the affinity labelled-biotin transport system in yeast cells.

Transport of biotin by Saccharomyces cerevisiae is inhibited by biotynyl p-nitrophenyl ester. Conversion of the inhibited cells to spheroplasts or simple treatment with thiols results in a total restoration of vitamin transport. Biotynyl p-nitrophenyl ester-induced inhibition is not due to an intracellular accumulation of the vitamin and consequent regulation, but appears to be due to specific labelling of the transport system.

Biological Transport↗

Identification of a region in 23S rRNA located at the peptidyl transferase center.

A photolyzable derivative of dipeptidyl-tRNA, p-azido-N-tBoc-Phe-[3H]Phe-tRNA, bound reversibly to 70S ribosomes in the presence of poly(U), becomes, when irradiated, covalently attached to components of the 50S ribosomal subunit. Most of the reaction occurs within the 23S RNA, while ribosomal proteins are only weakly labeled. Reversible binding as well as the covalent reaction are reduced in the absence of poly(U) or in the presence of several antibiotics specific for the 50S ribosomal subunit. There are apparently few sites (or perhaps a single site) of reaction on the 23S RNA that are exclusively located within the 2000 nucleotides from the 3 terminus of the molecule. Part of the 23S RNA within this region must therefore be closely associated with the peptidyl transferase center of the ribosome.

Binding Sites↗

Structure of a soluble super-active insulin is revealed by the nature of the complex between cyanogen-bromide-activated sepharose and amines.

Insulin-like material with elevated insulin specific acitivity is released from insulin-Sepharose in the presence of bovine-serum albumin. The mechanism of release and the chemical nature of this insulin-like material are revealed by the finding that amine-Sepharose is O-Sepharose-N-substituted isourea. Nucleophilic attack by amino groups releases N-1-N-2-disubstituted guanidines. Correspondingly, it is shown that the super-active insulin-like material is an N-1-N-2-disubstituted guanidine in which insulin and bovine-serum albumin are the substituents.

Amines↗

Distribution of end-to-end distances of oligopeptides in solution as estimated by energy transfer.

A homologous series of oligopeptides each containing at its ends a donor and an acceptor of electronic excitation energy was synthesized by the solid-phase method. N-5-(2-Hydroxyethyl)-L-glutamine was the repeating unit, and peptides containing 4, 5, 6, 7, 8, and 9 of these amino-acid residues were prepared. The chromophores naphthalene and dansyl, which were used as donor and acceptor, respectively, fulfil the conditions necessary for energy transfer according to the Förster mechanism. A distance corresponding to 50% efficiency of energy transfer, tro = 22 plus or minus 1 A, was calculated. The kinetics of fluorescence decay of an oligomer containing the naphthalene chromophore only could be described precisely by a monoexponential function. In contrast, the kinetics of the decay curves of the fluorescence of the donor of all of the oligomers containing both donor and acceptor, as measured in viscous solution, deviated markedly from monoexponential behavior. The deviation was interpreted in terms of the great number of different conformations that the various molecules of each of the oligomers attain in solution, leading to characteristic end-to-end distribution functions between the donor and acceptor. Numerical adjustment of the parameters of some of the previously proposed expressions to describe the end-to-end distribution enabled the reconstruction of the kinetics of the fluorescence decay of the donor with great precision. The end-to-end distribution functions for the various oligopeptides were thus evaluated.

Alanine↗

The covalent binding of daunomycin and adriamycin to antibodies, with retention of both drug and antibody activities.

Daunomycin and adriamycin, two potent cancer chemotherapeutic agents, were linked to immunoglobulins, making use of various covalent cross-linking methods. The most suitable method for binding of the drugs to the antibodies, which retained both antibody and drug activity, was periodate oxidation of the drug, followed by the linking of the oxidized drug to the immunoglobulin and subsequent reduction of the product with sodium borohydride. The activity of the drug-antibody conjugates was tested in vitro on tumor and normal cell cultures and was found to be similar to that of the free drug. A significant amount of antibody activity was retained, as found both with anti-bovine serum albumin antibodies, assayed by chemically modified bacteriophage, and with anti-mouse tumor antibodies, assayed by C'-dependent cytotoxicity.

Animals↗