Abstract
Fluorescence studies have been performed on the interaction of papain with active-site-directed inhibitors of the type mansyl-(Gly)n-Phe-glycinal, where n = 0, 1, 2. It has been found that whereas the mansyl [6-(N-methylantilino)-2-naphthalene sulfonyl] fluorescence of mansyl-Phe-glycinal is greatly enhanced, that of the two longer mansyl compounds is not, although all three are equally effective as inhibitors of papain action. Measurements of fluorescence polarization and rotational relaxation time support the conclusion that the fluorescent probe group of the two longer mansyl compounds protrudes into the solvent to a greater degree than that of mansyl-Phe-glycinal. Considerable energy transfer from papain tryptophan to the mansyl group is evident for all three inhibitors, however, although it is most marked with mansyl-Phe-glycinal. Stopped-flow fluorescence measurements have shown that, after initial rapid interaction, the first-order conformational changes in the active-site region of papain in the complex with mansyl-Phe-glycinal are approximately 1/10(4) those observed with comparable mansyl oligopeptide substrates, and approximately 1/10(2) those with acetyl-Phe-glycinal.
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Selected References
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- Bendall M. R., Cartwright I. L., Clark P. I., Lowe G., Nurse D. Inhibition of papain by N-acyl-aminoacetaldehydes and N-acyl-aminopropanones. Evidence for hemithioacetal formation by a cross-saturation technique in nuclear-magnetic resonance spectroscopy. Eur J Biochem. 1977 Sep 15;79(1):201–209. doi: 10.1111/j.1432-1033.1977.tb11798.x. [DOI] [PubMed] [Google Scholar]
- Fruton J. S. The mechanism of the catalytic action of pepsin and related acid proteinases. Adv Enzymol Relat Areas Mol Biol. 1976;44:1–36. doi: 10.1002/9780470122891.ch1. [DOI] [PubMed] [Google Scholar]
- Henderson P. J. A linear equation that describes the steady-state kinetics of enzymes and subcellular particles interacting with tightly bound inhibitors. Biochem J. 1972 Apr;127(2):321–333. doi: 10.1042/bj1270321. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis C. A., Jr, Wolfenden R. Thiohemiacetal formation by inhibitory aldehydes at the active site of papain. Biochemistry. 1977 Nov 1;16(22):4890–4895. doi: 10.1021/bi00641a023. [DOI] [PubMed] [Google Scholar]
- Lowe G., Whitworth A. S. A kinetic and fluorimetric investigation of papain modified at tryptophan-69 and -177 by N-bromosuccinimide. Biochem J. 1974 Aug;141(2):503–515. doi: 10.1042/bj1410503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mattis J. A., Fruton J. S. Kinetics of the action of papain on fluorescent peptide substrates. Biochemistry. 1976 May 18;15(10):2191–2194. doi: 10.1021/bi00655a025. [DOI] [PubMed] [Google Scholar]
- Mattis J. A., Henes J. B., Fruton J. S. Interaction of papain with derivatives of phenylalanylglycinal. J Biol Chem. 1977 Oct 10;252(19):6776–6782. [PubMed] [Google Scholar]
- Morgan G., Fruton J. S. Kinetics of the action of thermolysin on peptide substrates. Biochemistry. 1978 Aug 22;17(17):3562–3568. doi: 10.1021/bi00610a022. [DOI] [PubMed] [Google Scholar]
- Schechter I., Berger A. On the active site of proteases. 3. Mapping the active site of papain; specific peptide inhibitors of papain. Biochem Biophys Res Commun. 1968 Sep 6;32(5):898–902. doi: 10.1016/0006-291x(68)90326-4. [DOI] [PubMed] [Google Scholar]
- Westerik J. O., Wolfenden R. Aldehydes as inhibitors of papain. J Biol Chem. 1972 Dec 25;247(24):8195–8197. [PubMed] [Google Scholar]