A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver.

A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Research Abstract Details 

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A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Abstract Text:

M Takiguchi,M Mori,M Tatibana,

A procedure is described for the preparation of free and membrane-bound polysomes from rat liver. The procedure involves: differential centrifugation of liver homogenate to separate free and membrane-bound polysomes; treatment of the membrane-bound polysome fraction with a detergent to release bound polysomes from membranes; and magnesium precipitation of both classes of polysomes. Free and bound polysomes prepared in this manner were essentially undegraded and highly active in cell-free protein synthesis. The recovery of polysomes was nearly quantitative and the distribution between the free and membrane-bound state was 41 and 59%, respectively. Polypeptides synthesized in vitro by the free and membrane-bound polysomes were quite different. The majority (81-84%) of mRNA activities of two secretory proteins (albumin and transferrin) were recovered in the membrane-bound polysomes, whereas the majority (81-85%) of mRNA activities of two cytosolic [aldolase B, EC 4.1.2.13, and argininosuccinate synthetase, EC 6.3.4.5], one mitochondrial [ornithine carbamoyltransferase, EC 2.1.3.3] and one peroxisomal [catalase, EC 1.11.1.6] proteins were recovered in the free polysomes. A polysome class synthesizing ornithine carbamoyltransferase was purified 42-fold from the free polysomes by immunoprecipitation. The procedure is rapid (4-5 h) and reproducible, and provides a nearly quantitative means of separating the two classes of polysomes.

A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Publishing Authors By Initials

M Takiguchi,M Mori,M Tatibana,

For similar investigative techniques: centrifugation: ultracentrifugation research abstracts see: investigative techniques: centrifugation: ultracentrifugation research

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A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 97

Page Numbers: 1447-59

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: May

YEAR: 1985

A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Information

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LANGUAGE: eng

NlmUniqueID: 376600

A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Keywords Mesh Terms:

KEYWORDS: Ultracentrifugation

MESH TERMS: pharmacology

Chemical & Substance for Abstract: A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. Information

Substance Name: Potassium Chloride

Registry Number: 7447-40-7

Grant and Affiliation Information for A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver.

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Country: JAPAN

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MEDLINETA: J Biochem

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