Download e-book for kindle: Affinity Labeling: Affinity Labeling by Nathan P. Kaplan, William B. Jakoby, Meir Wilchek

By Nathan P. Kaplan, William B. Jakoby, Meir Wilchek

The seriously acclaimed laboratory typical, equipment in Enzymology, is likely one of the such a lot hugely revered guides within the box of biochemistry. considering the fact that 1955, every one quantity has been eagerly awaited, often consulted, and praised through researchers and reviewers alike. The sequence comprises a lot fabric nonetheless suitable this present day - really an important e-book for researchers in all fields of lifestyles sciences.

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Biochem. Biophys. 165, 739 (1974). [3] M e c h a n i s m - B a s e d I r r e v e r s i b l e Enzyme Inhibitors By ROBERTR. RANDO There are two extreme forms of mechanism for specific irreversible enzyme inhibitors. One comprises the classical affinity labeling agents, which are substrate analogs containing chemically reactive functional groups. 1 The specific binding of these inhibitors to the active-site regions of the enzyme ensures a greater probability of chemical reaction with a residue within this region as opposed to nonspecific chemical reactions.

T. Freer, and J. Kraut, J. Biol. Chem. 250, 7120 (1975). ~*K. A. Koehler and G. E. Lienhard, Biochemistry 10, 2477 (1971). de inhibitors that may form adducts resembling tetrahedral intermediates in substrate hydrolysis, vary markedly but in parallel as the structure of the substrate (or inhibitor) is varied at points distant from the scissile bondJ ~ Cases of this kind are especially interesting in that they seem to imply the occurrence of conformation changes in the enzyme and/or the substrate during the catalytic process.

As expected, fl, ~,-unsaturated a-ketoacids are irreversible inhibitors of the enzyme in the pyridoxamine form2 Finally, the pH rate profile for the inhibitor is the same as that for substrate turnoveff; this need not be the case, but does obtain in this example. 8 gM, some three orders of magnitude lower than the Km for the natural substrate, y-aminobutyrate. 3 gM gabaculine. As expected, gabaculine is not an irreversible inhibitor of the enzyme in the pyridoxamine form, and the mechanism of the irreversible inhibition by gabaculine is completely in accord with the hypothesis that enzymic conversion before inhibition.

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Affinity Labeling: Affinity Labeling by Nathan P. Kaplan, William B. Jakoby, Meir Wilchek

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