Low-pH association of proteins with the membranes of intact red blood cells. II. Studies of the mechanism.
The low-pH interaction of proteins with erythrocyte membranes has been found to be correlated with pH-induced changes in the erythrocyte membrane. Using a 90 degree lightscattering method it was shown that red blood cell hemolysis was slow between pH 5.8 and 5 (t1/2 above 1 h) but became fast at and below pH 4.7 (t1/2 less than 20 min). At pH 4.7, the presence of glycophorin in the incubation medium inhibited the hemolysis of erythrocytes and this protective effect was found to be dependent on the glycophorin concentration. Electron microscope experiments showed the presence of membrane defects after 10 s incubation at pH 4.6 in the absence of glycophorin in the incubation medium. These defects could further develop into openings with average widths of 14 nm after 1.5 min incubation under the acidic conditions. Fluorescence and flow cytometry studies showed that at pH 4.7, but not at pH 7.4, glycophorin tightly associates with phosphatidylcholine liposomes, and that liposome associated glycophorin molecules are recognized by anti-glycophorin monoclonal antibodies.
Arvinte
T
T
Biophor Corporation, Texas A & M University Research Park, College Station 77840.
Cudd
A
A
Schulz
B
B
Nicolau
C
C
D002450:Cell Communication; D004910:Erythrocyte Membrane / Q000502:physiology* / Q000648:ultrastructure; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005454:Fluorescence Polarization; D006021:Glycophorins / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008565:Membrane Proteins / Q000097:blood* / Q000502:physiology / Q000648:ultrastructure; D010713:Phosphatidylcholines / Q000502:physiology; D012542:Scattering, Radiation; D013861:Thiocyanates
D005452:Fluoresceins; D006021:Glycophorins; D008081:Liposomes; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate