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Hyaluronate-binding protein of simian virus 40-transformed 3T3 cells: Membrane distribution and reconstitution into lipid vesicles by Gloria Chi-Rosso; Bryan P. Toole is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Hyaluronate-binding protein (HABP) has been extracted in detergent from the membranes of simian virus 40-transformed 3T3 (SV-3T3) cells (Underhill et al, J Biol Chem 258:8086-8091, 1983). When SV-3T3 cells were treated with trypsin prior to isolation and dissolution of the membranes, no hyaluronate-binding activity could be detected. This indicates that all of the detectable HABP of SV-3T3 cells is located on the external surface of the plasma membrane rather than on internal membranes, which would be inaccessible to the trypsin. The detergentextracted HABP from SV-3T3 membranes was reconstituted into the membrane of lipid vesicles, which were formed by addition of exogenous phosphatidylcholine and cholic acid to the extracts followed by removal of detergent by dialysis against 0.02 M Tris pH 8.0 in the presence of protease inhibitors. Reconstitution was assessed by sedimentation in a discontinuous sucrose gradient and by gel filtration on Sepharose 4B in the presence and absence of detergent. The characteristics of binding of hyaluronate to the reconstituted HABP were then compared with those studied previously for the original membrane-bound HABP and the detergentextracted HABP (

Who reads Hyaluronate-binding protein of simian virus 40-transformed 3T3 cells: Membrane distribution and reconstitution into lipid vesicles?

Author

Gloria Chi-Rosso; Bryan P. Toole

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 0730-2312)

Published

1987

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)