Rotational mobility of the fibrinogen receptor glycoprotein IIb/IIIa or integrin alpha IIb beta 3 in the plasma membrane of human platelets.
Integrin alpha IIb beta 3 or glycoprotein IIb/IIIa (GPIIb/IIIa, 228 kDa) is a Ca(2+)-dependent, noncovalent heterodimer of glycoproteins IIb (GPIIb or alpha IIb, 136 kDa) and IIIa (GPIIIa or beta 3, 92 kDa), which serves as the receptor for fibrinogen and other adhesive proteins at the surface of activated platelets. We have determined the microsecond-range rotational motions of alpha IIb beta 3 in resting platelets, in isolated plasma membranes, and reconstituted in 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) bilayers. The measurements were based on the time-resolved phosphorescence anisotropy [r(t)] of erythrosin-labeled F(ab) fragments [Er-F(ab)] of monoclonal antibodies bound to alpha IIb beta 3. In general, the r(t) decays were satisfactorily fitted to the sum of the two exponential terms and a constant, from which the initial anisotropy (r(in) approximately 0.05-0.11), the short (phi 1 approximately 1-14 microseconds) and the long (phi 2 approximately 15-60 microseconds) rotational correlation times, and the limiting anisotropy (r infinity approximately 0.02-0.07) were obtained. The observed values depended on the platelet preparation, temperature, Ca2+ concentration, and the antibody used. In accordance with data on the order parameter and the viscosity of the lipid bilayer of the platelet plasma membrane, phi 2 and r infinity of the alpha IIb beta 3-Er-F(ab) complexes in the three preparations decreased with the increase of temperature, the r(t) curves being fully reversible within the interval from 5 to 35 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)
González-Rodríguez
J
J
Unidad de Biofísica, Instituto de Química Física, CSIC, Madrid, Spain.
Acuña
A U
AU
Alvarez
M V
MV
Jovin
T M
TM
D000911:Antibodies, Monoclonal; D001665:Binding Sites; D001666:Binding Sites, Antibody; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000097:blood; D002462:Cell Membrane / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism; D005454:Fluorescence Polarization; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D016023:Integrins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008163:Luminescent Measurements; D008961:Models, Structural; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D013816:Thermodynamics; D013997:Time Factors
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D016023:Integrins; D008081:Liposomes; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; D005340:Fibrinogen; D004492:Edetic Acid; D002118:Calcium