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Title: Optical probe for the cytochrome P-450 cholesterol side chain cleavage enzyme

Abstract

An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450.sub.scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450.sub.scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.

Inventors:
 [1];  [1];  [1];  [2]
  1. Los Alamos, NM
  2. Santa Fe, NM
Issue Date:
Research Org.:
Los Alamos National Laboratory (LANL), Los Alamos, NM
OSTI Identifier:
868276
Patent Number(s):
5110725
Assignee:
United States of America as represented by United States (Washington, DC)
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
optical; probe; cytochrome; p-450; cholesterol; chain; cleavage; enzyme; enables; study; activity; absorbance; spectroscopy; sensitive; fluorescence; methods; particular; provides; ability; monitor; scc; rate; limiting; steroid; biosynthesis; located; inner; mitochondrial; membrane; catalyzes; conversion; pregnenolone; isocapraldehyde; sequential; oxidations; fluorogenic; cholesterol-like; linked; chromophore; linking; selected; response; substrate; enhanced; cleaved; fluorescent; anion; optically; detected; generated; side-chain; reaction; steroidogenesis; enzyme activity; optical response; optical probe; optically detected; cytochrome p-450; probe provides; sensitive fluorescence; chain cleavage; cleavage enzyme; p-450 cholesterol; fluorescence method; fluorescence methods; probe enables; absorbance spectroscopy; /435/

Citation Formats

Marrone, Babetta L, Simpson, Daniel J, Unkefer, Clifford J, and Whaley, Thomas W. Optical probe for the cytochrome P-450 cholesterol side chain cleavage enzyme. United States: N. p., 1992. Web.
Marrone, Babetta L, Simpson, Daniel J, Unkefer, Clifford J, & Whaley, Thomas W. Optical probe for the cytochrome P-450 cholesterol side chain cleavage enzyme. United States.
Marrone, Babetta L, Simpson, Daniel J, Unkefer, Clifford J, and Whaley, Thomas W. Wed . "Optical probe for the cytochrome P-450 cholesterol side chain cleavage enzyme". United States. https://www.osti.gov/servlets/purl/868276.
@article{osti_868276,
title = {Optical probe for the cytochrome P-450 cholesterol side chain cleavage enzyme},
author = {Marrone, Babetta L and Simpson, Daniel J and Unkefer, Clifford J and Whaley, Thomas W},
abstractNote = {An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450.sub.scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450.sub.scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1992},
month = {1}
}

Patent:

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