SINGLE CELL ENDOCRINOLOGY - ANALYSIS OF P-450SCC ACTIVITY BY FLUORESCENCE DETECTION METHODS

被引：13

作者：

MARRONE, BL ^{[1
]}

SIMPSON, DJ ^{[1
]}

YOSHIDA, TM ^{[1
]}

UNKEFER, CJ ^{[1
]}

WHALEY, TW ^{[1
]}

BUICAN, TN ^{[1
]}

机构：

[1] UNIV CALIF LOS ALAMOS SCI LAB,DIV ISOTOPE & NUCL CHEM,LOS ALAMOS,NM 87545

来源：

ENDOCRINOLOGY | 1991年 / 128卷 / 05期

关键词：

D O I：

10.1210/endo-128-5-2654

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

A mechanism-based, fluorogenic probe for the cytochrome P-450scc (cholesterol side chain cleavage) enzyme, rate-limiting for the conversion of cholesterol to steroid hormones, is introduced and its application to the study of enzyme activity and regulation in single steroidogenic cells by several fluorescence detection methods is demonstrated. Reaction of the probe with P-450scc gives pregnenolone and the highly fluorescent resorufin anion. Spectroscopic changes in probe fluorescence, indicative of P-450scc activity, were monitored by steady-state fluorescence spectroscopy, flow cytometry, and microspectrofluorometry. This unique probe provides a nonradiometric indicator for real-time measurement of P-450scc activity in single living cells.

引用

页码：2654 / 2656

页数：3

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