Synthesis, processing, and intracellular transport of CD36 during monocytic differentiation.
CD36 is an integral membrane glycoprotein expressed by several cell types, including endothelial cells of the microvasculature, erythrocytes, platelets, and monocytes. In the monocytic lineage, CD36 is expressed during the late stages of differentiation in the bone marrow, in circulating monocytes, and in some tissue resident macrophages, and it is thought to mediate the phagocytosis of apoptotic cells and the endocytic uptake of modified lipoproteins. Here we analyze the synthesis, processing, and intracellular transport of CD36 in U937 and THP-1, two human cell lines representing different stages of monocytic maturation. In both cell lines, phorbol 12-myristate 13-acetate induces the expression of CD36. A 74-kDa intracellular precursor is first synthesized that has the hallmarks of a resident protein of the endoplasmic reticulum. The precursor protein is later processed into a mature form of 90-105 kDa which is transported to the cell surface. The kinetics of processing differ significantly in U937 and THP-1. These differences are specific for the CD36, as two unrelated proteins (CD11b and CD45R) are processed and transported to the surface at similar rates in the two cell lines. A 33-kDa endoglycosidase H-sensitive glycoprotein specifically associates with the 74-kDa precursor. Coprecipitation of gp33 correlates with slow processing of CD36 precursor, suggesting that gp33 may play a role in regulating the intracellular transport of CD36, during monocyte maturation.
Alessio
M
M
DIBIT, San Raffaele Scientific Institute, Milan, Italy.
De Monte
L
L
Scirea
A
A
Gruarin
P
P
Tandon
N N
NN
Sitia
R
R
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D015153:Blotting, Western; D018955:CD36 Antigens / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D006026:Glycoside Hydrolases; D018922:HL-60 Cells; D006801:Humans; D007700:Kinetics; D007938:Leukemia; D009000:Monocytes / Q000166:cytology / Q000276:immunology*; D011499:Protein Processing, Post-Translational; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018955:CD36 Antigens; D006026:Glycoside Hydrolases; D013755:Tetradecanoylphorbol Acetate