A CONTINUOUS SPECTROPHOTOMETRIC DIRECT ASSAY FOR PEPTIDYL PROLYL CIS-TRANS-ISOMERASES

被引：23

作者：

GARCIAECHEVERRIA, C ^{[1
]}

KOFRON, JL ^{[1
]}

KUZMIC, P ^{[1
]}

RICH, DH ^{[1
]}

机构：

[1] UNIV WISCONSIN,DEPT CHEM,MADISON,WI 53706

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1993年 / 191卷 / 01期

关键词：

D O I：

10.1006/bbrc.1993.1185

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

M-Nitrotyrosine incorporated into proline peptides of the general sequence -Xxx-Pro-Tyr(m-NO2)- responds to cis-trans Xxx-Pro conformational transition by changes in the pK(a) of its side-chain hydroxyl (Garel and Siffert, 1979). We exploited this effect to develop a continuous direct (uncoupled) assay for peptidyl prolyl cis-trans isomerases. Prior to the enzyme assay, the cis-trans equilibrium is perturbed in favor of the cis isomer by dissolving the substrate H-Ala-Ala-Pro-Tyr(m-NO2)-Ala-NH2 in a 470 mM solution of LiCl in trifluoroethanol. Upon addition of substrate to the biological buffer, the conformational equilibrium characteristic for the aqueous medium is restored, and the Ala-Pro isomerization is monitored spectrophotometrically. © 1993 Academic Press, Inc.

引用

页码：70 / 75

页数：6

共 38 条

[1] PREPARATION AND APPLICATION OF THE 5-(4-(9-FLUORENYLMETHYLOXYCARBONYL)AMINOMETHYL-3,5-DIMETHOXYPHENOXY)VALERIC ACID (PAL) HANDLE FOR THE SOLID-PHASE SYNTHESIS OF C-TERMINAL PEPTIDE AMIDES UNDER MILD CONDITIONS [J].