Identification of two lysosomal membrane glycoproteins.
Two murine lysosome-associated membrane proteins, LAMP-1 of 105,000-115,000 D and LAMP-2 of 100,000-110,000 D, have been identified by monoclonal antibodies that bind specifically to lysosomal membranes. Both glycoproteins were distinguished as integral membrane components solubilized by detergent solutions but not by various chaotropic agents. The lysosome localization was demonstrated by indirect immunofluorescent staining, co-localization of the antigen to sites of acridine orange uptake, and immunoelectron microscopy. Antibody binding was predominantly located at the limiting lysosomal membrane, distinctly separated from colloidal gold-labeled alpha-2-macroglobulin accumulated in the lumen during prolonged incubation. LAMP-1 and LAMP-2 also appeared to be present in low concentrations on Golgi trans-elements but were not detected in receptosomes marked by the presence of newly endocytosed alpha-2-macroglobulin, or in other cellular structures. LAMP-1 and LAMP-2 were distinguished as different molecules by two-dimensional gel analysis, 125I-tryptic peptide mapping, and sequential immunoprecipitations of 125I-labeled cell extracts. Both glycoproteins were synthesized as a precursor protein of approximately 90,000 D, and showed a marked heterogeneity of apparent molecular weight expression in different cell lines. LAMP-2 was closely related or identical to the macrophage antigen, MAC-3, as indicated by antibody adsorption and tryptic peptide mapping. It is postulated that these glycoproteins, as major protein constituents of the lysosomal membrane, have important roles in lysosomal structure and function.
J W
JW
Chen
T L
TL
Murphy
M C
MC
Willingham
I
I
Pastan
J T
JT
August
D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D000954:Antigens, Surface / Q000032:analysis; D002460:Cell Line; D011992:Endosomes / Q000378:metabolism; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D007425:Intracellular Membranes / Q000032:analysis; D007526:Isoelectric Point; D051907:Lysosomal Membrane Proteins; D008247:Lysosomes / Q000032:analysis*; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D000511:alpha-Macroglobulins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D000954:Antigens, Surface; D006023:Glycoproteins; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; D010446:Peptide Fragments; D000511:alpha-Macroglobulins; C036763:monocyte-macrophage differentiation antigen