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

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

The significantly acclaimed laboratory normal, equipment in Enzymology, is likely one of the such a lot hugely revered courses within the box of biochemistry. considering 1955, every one quantity has been eagerly awaited, often consulted, and praised through researchers and reviewers alike. The sequence includes a lot fabric nonetheless correct at the present time - actually a vital ebook for researchers in all fields of existence sciences.

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E. W. Miles, Biochem. Biophys. Res. Commun. 66, 94 (1975). 32 ~] GENERAL METHODOLOGY tryptophan synthetase by 2, and ~-cystathionase by 3. The inhibition of aspartate aminotransferase by 1 and ~ involves the sequence of steps shown in Eq. (4). j *~N/°~. ~ H H (4) when R--OCH3 (4) -- OCH~ Enz ~ e ~ e f C O 2" CHa I I ÷ II tt Since both inhibitors must be converted into the active form 4 by the enzyme, neither one inhibits the holoenzyme in the pyridoxamine form or the apoenzyme. 5 The reactive enzyme product itself must be an affinity labeling agent of the enzyme.

Both of the latter reagents are first turned over by the enzyme before inhibition ensues, but the mechanisms of inhibition remain obscure. Recently, N-nitroso compounds have been introduced as irreversible inhibitors of proteolytie enzymes. 27 R. R. Rando, J. Am. Chem. Soc. 95, 4438 (1973). 2~R. C. ttevey, J. Babson, A. L. Maycock, and R. Abeles, J. Am. Chem. Soc. 95, 6125 (1973). ~B. T. Ho, J. Pharm. Sci. 61, 821 (1972). E. I=l. White, D. F. Roswell, I. R. Politzer, and B. R. Branchini, J. Am.

Strachan, and G. A. Levvy, Biochem. J. 102, 929 (1967). t It. L. Lai and B. Axelrod, Biochem. Biophys. Res. Commun. 64, 463 (1973). B. E. Evans and R. Wolfenden, J. Am. Chem. Soc. 92, 4751 (1970). P. W. K. Woo, It. W. Dion, S. M. Lange, L. F. Dahl, and L. J. Durham, J. Helerocycl. Chem. 11, 641 (1974). R. M. Cohen and R. Wolfenden, J. Biol. Chem. 246, 7561 (1971). L. D. Byers and R. Wolfenden, J. Biol. Chem. 247, 606 (1972). A. Schmitt, I. Bottke, and G. Siebert, Hoppe-Seyler's Z. Physiol. Chem.

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