CHARACTERIZATION OF PROTEASE-CATALYZED HYDROLYSIS OF CYANINE-LABELED ANGIOTENSIN USING CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION

被引:9
作者
CHEN, FTA
机构
[1] Advanced Technology Center, Beckman Instruments Inc., Fullerton, CA 92634, 2500 Harbor Boulevard
关键词
D O I
10.1006/abio.1995.1164
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The hydrolytic cleavage of a cyanine (Cy3)-labeled angiotensin, catalyzed by various proteases, was studied by capillary electrophoresis (CE) with laser-induced fluorescence detection (LIF). The end-labeled peptides and the Cy3 diacid internal standard were separated on a 20-mu m X 27-cm capillary with LIF detection (emission, 580 nm) using a frequency-doubled solid-state diode laser emitting at 532 nm or a He-Ne laser emitting at 543 nm. Hydrolysis of the Cy3-labeled angiotensin I, catalyzed by proteinase K, is a sequential process beginning from the C-terminal of the peptide, instead of from random cleavages. Trypsin catalyzes a specific cleavage of Cy3-angiotensin I to Cy3-Asp-Arg as anticipated, Using a combination of endopeptidase and carboxypeptidases, the remnant of the labeled species was characterized by CE-LIF. The method provides a general tool for studying the mechanism of protease-catalyzed hydrolysis of peptide. (C) 1995 Academic Press, Inc.
引用
收藏
页码:341 / 345
页数:5
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