The intracellular localization of adrenal 3β-hydroxysteroiddehydrogenase/Δ5-isomerase by density gradient perturbation

Denis R. Headon; Joyce Hsiao; Frank Ungar

doi:10.1016/0006-291X(78)90883-5

The intracellular localization of adrenal 3β-hydroxysteroiddehydrogenase/Δ⁵-isomerase by density gradient perturbation

Denis R. Headon
, Joyce Hsiao
, Frank Ungar

University of Minnesota Medical School

Research output: Contribution to a Journal (Peer & Non Peer) › Article › peer-review

12 Citations (Scopus)

Abstract

The intracellular localization of 3β-hydroxysteroiddehydrogenase/Δ⁵-isomerase (Δ⁵-3β-OHD) in bovine adrenal cortex was determined by density perturbation with density gradient centrifugation and the use of marker enzymes. Mitochondrial rich fractions after incubation with succinate and iodonitrotetrazolium (INT) were loaded with insoluble reduced formazan and separated on a linear sucrose gradient. A shift in median density was observed in the INT-treated mitochondria for protein, succinic dehydrogenase activity, and for Δ⁵-3β-OHD activity, but not for 21-hydroxylase, a microsomal enzyme marker. Density perturbation and gradient fractionation reveals a dual location of Δ⁵-3β-OHD in both mitochondrial and microsomal fractions.

Original language	English
Pages (from-to)	1006-1012
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	82
Issue number	3
DOIs	https://doi.org/10.1016/0006-291X(78)90883-5
Publication status	Published - 14 Jun 1978
Externally published	Yes

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title = "The intracellular localization of adrenal 3β-hydroxysteroiddehydrogenase/Δ5-isomerase by density gradient perturbation",

abstract = "The intracellular localization of 3β-hydroxysteroiddehydrogenase/Δ5-isomerase (Δ5-3β-OHD) in bovine adrenal cortex was determined by density perturbation with density gradient centrifugation and the use of marker enzymes. Mitochondrial rich fractions after incubation with succinate and iodonitrotetrazolium (INT) were loaded with insoluble reduced formazan and separated on a linear sucrose gradient. A shift in median density was observed in the INT-treated mitochondria for protein, succinic dehydrogenase activity, and for Δ5-3β-OHD activity, but not for 21-hydroxylase, a microsomal enzyme marker. Density perturbation and gradient fractionation reveals a dual location of Δ5-3β-OHD in both mitochondrial and microsomal fractions.",

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year = "1978",

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doi = "10.1016/0006-291X(78)90883-5",

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T1 - The intracellular localization of adrenal 3β-hydroxysteroiddehydrogenase/Δ5-isomerase by density gradient perturbation

AU - Headon, Denis R.

AU - Hsiao, Joyce

AU - Ungar, Frank

PY - 1978/6/14

Y1 - 1978/6/14

N2 - The intracellular localization of 3β-hydroxysteroiddehydrogenase/Δ5-isomerase (Δ5-3β-OHD) in bovine adrenal cortex was determined by density perturbation with density gradient centrifugation and the use of marker enzymes. Mitochondrial rich fractions after incubation with succinate and iodonitrotetrazolium (INT) were loaded with insoluble reduced formazan and separated on a linear sucrose gradient. A shift in median density was observed in the INT-treated mitochondria for protein, succinic dehydrogenase activity, and for Δ5-3β-OHD activity, but not for 21-hydroxylase, a microsomal enzyme marker. Density perturbation and gradient fractionation reveals a dual location of Δ5-3β-OHD in both mitochondrial and microsomal fractions.

AB - The intracellular localization of 3β-hydroxysteroiddehydrogenase/Δ5-isomerase (Δ5-3β-OHD) in bovine adrenal cortex was determined by density perturbation with density gradient centrifugation and the use of marker enzymes. Mitochondrial rich fractions after incubation with succinate and iodonitrotetrazolium (INT) were loaded with insoluble reduced formazan and separated on a linear sucrose gradient. A shift in median density was observed in the INT-treated mitochondria for protein, succinic dehydrogenase activity, and for Δ5-3β-OHD activity, but not for 21-hydroxylase, a microsomal enzyme marker. Density perturbation and gradient fractionation reveals a dual location of Δ5-3β-OHD in both mitochondrial and microsomal fractions.

UR - https://www.scopus.com/pages/publications/0018138882

U2 - 10.1016/0006-291X(78)90883-5

DO - 10.1016/0006-291X(78)90883-5

M3 - Article

C2 - 567983

AN - SCOPUS:0018138882

SN - 0006-291X

VL - 82

SP - 1006

EP - 1012

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 3

ER -

The intracellular localization of adrenal 3β-hydroxysteroiddehydrogenase/Δ⁵-isomerase by density gradient perturbation

Abstract

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