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The Journal of Cell Biology, Vol 47, 34-48, Copyright © 1970 by Rockefeller University Press

ARTICLE

SUBFRACTIONATION OF SMOOTH MICROSOMES FROM RAT LIVER

Hans Glaumann 1 and Gustav Dallner 1

1 From the Department of Pathology at Sabbatsberg Hospital, Karolinska Institutet, Stockholm, Sweden

Total smooth microsomes from rat liver isolated on a Cs+-containing sucrose gradient were concentrated and subsequently fractionated by zone centrifugation on a stabilizing sucrose gradient. The prerequisite for fractionation is to prepare total smooth microsomes in a nonaggregated condition, as well as to utilize a procedure which counteracts enzyme inactivation. The median equilibrium density of the various smooth microsomal vesicles ranges from 1.10 to 1.18. The phospholipid/protein ratio is identical in all subfractions, but cholesterol, on a PLP basis, is enriched in the subfractions with the highest sedimentation velocity. The enzyme distribution pattern reveals a pronounced heterogeneity. A number of NADH- and NADPH-oxidizing enzymes are concentrated in the upper part of the gradient and exhibit a certain degree of separation from G6Pase. Mg++-ATPase and AMPase are enriched in the lower part of the gradient. No specific enrichment of newly synthesized NADPH-cytochrome c reductase activity occurs in any of the subfractions after phenobarbital treatment. These data demonstrate that smooth microsomes, by adequate fractionation procedure, can be separated into subfractious of heterogeneous composition.

Submitted on October 16, 1969
Revised on April 24, 1970


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