ISSN:
0730-2312
Keywords:
electron microscopy
;
plasma membrane
;
lymphoma cells
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
Notes:
Plasma membrane was isolated from the mouse T lymphoma cell line WEHI-22 using four different methods of cell disruption followed by centrifugal fractionation. Disruption by nitrogen cavitation or by shearing with a cell pump produced plasma membrane vesicles of similar buoyant density (1.10 g/ml) and morphological appearance. Few C-type virus particles were present. Cell disruption with 2% Tween-40 produced membrane vesicles of similar morphology but lower density (1.09 g/ml). All of the above preparations resulted in vesicles with aggregated intramembranous particles after freeze fracture. Microvesiculation with a sublytic concentration of a lysophosphatidylcholine analog (ET-12-H) (0.0032% w/v) produced small membrane vesicles which could be isolated without differential centrifugation. However, these had a slightly higher density than vesicles prepared by cavitation or shearing and were co ntaminated by virus particles. Unlike the other preparations, vesicles prepared with ET-I2-H had dispersed intramembranous particles. The enzyme γ-glutamyl transferase was enriched from 20- to 45-fold in the membrane preparations and proved a suitable plasma membrane marker for these cells whose 5′-nucleotidase content is very low.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/jcb.240200205
