ISOLATION OF SMOOTH VESICLES AND FREE RIBOSOMES FROM RAT LIVER MICROSOMES

Autor:	J. Chauveau, Yvonne Moulé, Ch. Rouiller, J. Schneebeli
Rok vydání:	1962
Předmět:	Differential centrifugation Membranes Vesicle RNA Cell Biology Biology Ribosome Article Rats Nucleoprotein Ferritin Nucleoproteins Membrane Biochemistry Microsomes Microsomes Liver biology.protein Microsome Animals Ribosomes Phospholipids
Zdroj:	The Journal of Cell Biology
ISSN:	1540-8140 0021-9525
DOI:	10.1083/jcb.12.1.17
Popis:	Microsomes, isolated from rat liver homogenate in 0.88 M sucrose, have been fractionated by differential centrifugation. The 2nd microsomal fraction, sedimented between 60 minutes at 105,000 g and 3 hours at 145,000 g, consists mainly of smooth vesicles, free ribosomes, and ferritin. By utilizing the differences in density existing between the membranes and the granular elements it has been possible to separate the smooth membranes from the free ribosomes and ferritin. The procedure is to resuspend the 2nd microsomal fraction in a sucrose solution of 1.21 or 1.25 density and centrifuge it at 145,000 g for 20 or 40 hours. A centripetal migration of membranes and a centrifugal sedimentation of granular elements are obtained. Phospholipids, as well as the enzymatic activities DPNH-cytochrome c reductase, glucose-6-phosphatase and esterase are localized in the membranes. The free ribosomes have been purified by washing. A concentration of 200 µg RNA per mg nitrogen has been reached. RNA is also present in the membranes. These results are discussed in relation to current views on microsomal structure and chemistry.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fe1a1cdc2d5d3b6e1d995f48688c57af https://doi.org/10.1083/jcb.12.1.17 Zobrazit plný text záznamu