Preparation and characterization of total,rough and smooth microsomes from the lung of control and methylcholanthrene-treated rats |
| |
Authors: | Karin Johannesen Joseph W Depierre Anders Bergstrand Gustav Dallner Lars Ernster |
| |
Institution: | 1. Arrhenius Laboratory, Department of Biochemistry, University of Stockholm, Fack, 104 05 Stockholm Sweden;2. Department of Pathology at Sabbatsberg Hospital, Karolinska Institutet, Stockholm Sweden |
| |
Abstract: | Optimal conditions for the preparation of relatively pure microsomes and microsomal subfractions from rat lung have been determined. The most important of these conditions is homogenization of a 20% (w/v) suspension of lung tissue in 0.44 M sucrose/1% (w/v) bovine serum albumin with four up-and-down strokes at 440 rev./min in a Potter-Elvehjem homogenizer. The 10 000 × g supernatant prepared from this homogenate can be centrifuged at 105 000 × g to obtain total microsomes or subfractionated into rough and smooth microsomes on a Cs+-containing discontinuous sucrose gradient. The total, rough and smooth microsomes have been characterized in terms of their chemical composition, enzymatic activity, and morphology. These preparations should prove useful in studies of various enzymes in lung (e.g. benzpyrene monooxygenase, epoxide hydrase, enzymes of phospholipid and ascorbic acid synthesis) and in subfractionations designed to reveal heterogeneites in the lateral plane of the lung endoplasmic reticulum. |
| |
Keywords: | AMPase adenosine 5′-monophosphatase IDPase inosine 5′-diphosphatase ATPase adenosine 5′-triphosphatase |
本文献已被 ScienceDirect 等数据库收录! |
|