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ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients

Rat liver microsomal fractions have been equilibrated in various types of linear density gradients. 15 fractions were collected and assayed for 27 constituents. As a result of this analysis microsomal constituents have been classified, in the order of increasing median density, into four groups labe...

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Autores principales: Beaufay, Henri, Amar-Costesec, Alain, Thinès-Sempoux, Denise, Wibo, Maurice, Robbi, Mariette, Berthet, Jacques
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1974
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2109260/
https://www.ncbi.nlm.nih.gov/pubmed/4150490
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author Beaufay, Henri
Amar-Costesec, Alain
Thinès-Sempoux, Denise
Wibo, Maurice
Robbi, Mariette
Berthet, Jacques
author_facet Beaufay, Henri
Amar-Costesec, Alain
Thinès-Sempoux, Denise
Wibo, Maurice
Robbi, Mariette
Berthet, Jacques
author_sort Beaufay, Henri
collection PubMed
description Rat liver microsomal fractions have been equilibrated in various types of linear density gradients. 15 fractions were collected and assayed for 27 constituents. As a result of this analysis microsomal constituents have been classified, in the order of increasing median density, into four groups labeled a, b, c, and d. Group a includes: monoamine oxidase, galactosyltransferase, 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and cholesterol; group b: NADH cytochrome c reductase, NADPH cytochrome c reductase, aminopyrine demethylase, cytochrome b (5), and cytochrome P 450; group c: glucose 6-phosphatase, nucleoside diphosphatase, esterase, β-glucuronidase, and glucuronyltransferase; group d: RNA, membrane-bound ribosomes, and some enzymes probably adsorbed on ribosomes: fumarase, aldolase, and glutamine synthetase. Analysis of the microsomal fraction by differential centrifugation in density gradient has further dissociated group a into constituents which sediment more slowly (monoamine oxidase and galactosyltransferase) than those of groups b and c, and 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and the bulk of cholesterol which sediment more rapidly (group a2). The microsomal monoamine oxidase is attributed, at least partially, to detached fragments of external mitochondrial membrane. Galactosyltransferase belongs to the Golgi complex. Group a2 constituents are related to plasma membranes. Constituents of groups b and c and RNA belong to microsomal vesicles derived from the endoplasmic reticulum. These latter exhibit a noticeable biochemical heterogeneity and represent at the most 80% of microsomal protein, the rest being accounted for by particles bearing the constituents of groups a and some contaminating mitochondria, lysosomes, and peroxisomes. Attention is called to the operational meaning of microsomal subfractions and to their cytological complexity.
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spelling pubmed-21092602008-05-01 ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients Beaufay, Henri Amar-Costesec, Alain Thinès-Sempoux, Denise Wibo, Maurice Robbi, Mariette Berthet, Jacques J Cell Biol Article Rat liver microsomal fractions have been equilibrated in various types of linear density gradients. 15 fractions were collected and assayed for 27 constituents. As a result of this analysis microsomal constituents have been classified, in the order of increasing median density, into four groups labeled a, b, c, and d. Group a includes: monoamine oxidase, galactosyltransferase, 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and cholesterol; group b: NADH cytochrome c reductase, NADPH cytochrome c reductase, aminopyrine demethylase, cytochrome b (5), and cytochrome P 450; group c: glucose 6-phosphatase, nucleoside diphosphatase, esterase, β-glucuronidase, and glucuronyltransferase; group d: RNA, membrane-bound ribosomes, and some enzymes probably adsorbed on ribosomes: fumarase, aldolase, and glutamine synthetase. Analysis of the microsomal fraction by differential centrifugation in density gradient has further dissociated group a into constituents which sediment more slowly (monoamine oxidase and galactosyltransferase) than those of groups b and c, and 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and the bulk of cholesterol which sediment more rapidly (group a2). The microsomal monoamine oxidase is attributed, at least partially, to detached fragments of external mitochondrial membrane. Galactosyltransferase belongs to the Golgi complex. Group a2 constituents are related to plasma membranes. Constituents of groups b and c and RNA belong to microsomal vesicles derived from the endoplasmic reticulum. These latter exhibit a noticeable biochemical heterogeneity and represent at the most 80% of microsomal protein, the rest being accounted for by particles bearing the constituents of groups a and some contaminating mitochondria, lysosomes, and peroxisomes. Attention is called to the operational meaning of microsomal subfractions and to their cytological complexity. The Rockefeller University Press 1974-04-01 /pmc/articles/PMC2109260/ /pubmed/4150490 Text en Copyright © 1974 by The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Article
Beaufay, Henri
Amar-Costesec, Alain
Thinès-Sempoux, Denise
Wibo, Maurice
Robbi, Mariette
Berthet, Jacques
ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title_full ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title_fullStr ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title_full_unstemmed ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title_short ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER : III. Subfractionation of the Microsomal Fraction by Isopycnic and Differential Centrifugation in Density Gradients
title_sort analytical study of microsomes and isolated subcellular membranes from rat liver : iii. subfractionation of the microsomal fraction by isopycnic and differential centrifugation in density gradients
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2109260/
https://www.ncbi.nlm.nih.gov/pubmed/4150490
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