Glycophorin as a receptor for Escherichia coli alpha-hemolysin in erythrocytes.
Escherichia coli alpha-hemolysin (HlyA) can lyse both red blood cells (RBC) and liposomes. However, the cells are lysed at HlyA concentrations 1-2 orders of magnitude lower than liposomes (large unilamellar vesicles). Treatment of RBC with trypsin, but not with chymotrypsin, reduces the sensitivity of RBC toward HlyA to the level of the liposomes. Since glycophorin, one of the main proteins in the RBC surface, can be hydrolyzed by trypsin much more readily than by chymotrypsin, the possibility was tested of a specific binding of HlyA to glycophorin. With this purpose, a number of experiments were performed. (a) HlyA was preincubated with purified glycophorin, after which it was found to be inactive against both RBC and liposomes. (b) Treatment of RBC with an anti-glycophorin antibody protected the cells against HlyA lysis. (c) Immobilized HlyA was able to bind glycophorin present in a detergent lysate of RBC ghosts. (d) Incorporation of glycophorin into pure phosphatidylcholine liposomes increased notoriously the sensitivity of the vesicles toward HlyA. (e) Treatment of the glycophorin-containing liposomes with trypsin reverted the vesicles to their original low sensitivity. The above results are interpreted in terms of glycophorin acting as a receptor for HlyA in RBC. The binding constant of HlyA for glycophorin was estimated, in RBC at sublytic HlyA concentrations, to be 1.5 x 10(-9) m.
Cortajarena
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Unidad de Biofisica (Consejo Superior de Investigaciones Cientificas-UPV/EHU), and Departamento de Bioquimica, Universidad del Pais Vasco/Euskal Herriko Unibertsitatea, Aptdo. 644, Bilbao 48080, Spain.
Goñi
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FM
Ostolaza
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D000818:Animals; D001426:Bacterial Proteins / Q000097:blood* / Q000737:chemistry; D001427:Bacterial Toxins / Q000097:blood* / Q000737:chemistry; D001665:Binding Sites; D004910:Erythrocyte Membrane / Q000502:physiology*; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006021:Glycophorins / Q000737:chemistry / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000097:blood* / Q000737:chemistry; D006736:Horses; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000097:blood; D014357:Trypsin
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D006021:Glycophorins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D008051:Lipid Bilayers; D011510:Proteolipids; D011956:Receptors, Cell Surface; C020485:proteoliposomes; D014357:Trypsin