Biochemical characterization by two-dimensional electrophoresis of lymphocyte antigens involved in cell-to-cell or cell-to-matrix adhesion.
We have exemplified three cases of application of two-dimensional (2-D) electrophoresis to the characterization of lymphocyte membrane antigens. We could show that the proteins recognized by two monoclonal antibodies, LAK1 and LAK2, on the surface of large granular cells mediating natural- and lymphokine-activated killing are distinct molecules. LAK1 is expressed without any structural modification, even on the surface of endothelial cells. Another membrane antigen, recognized by the monoclonal antibody FB12, was shown to have the overall structure of the integrins of the very late activation (VLA) class, being composed of an alpha and of a beta subunit. The latter corresponded to the beta 1 type as already characterized for other VLAs, whereas the alpha chain was different from alpha 1 through alpha 6. The 2-D protocol using immobilized pH gradients for the first dimension allows reliable assessment of the identity of individual components because of the reproducibility of the absolute coordinates for spot position.
Istituto Scientifico San Raffaele, Milano, Italy.
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Zocchi
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Fabbri
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Poggi
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Gianazza
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology* / Q000502:physiology; D002448:Cell Adhesion; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004730:Endothelium, Vascular / Q000502:physiology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D016023:Integrins / Q000032:analysis*; D011947:Receptors, Antigen, B-Cell / Q000032:analysis*; D013601:T-Lymphocytes / Q000276:immunology* / Q000502:physiology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D016023:Integrins; D011947:Receptors, Antigen, B-Cell