Formation of one or more intrachain disulphide bonds is required for the intracellular processing and transport of CD36.
In monocytes/macrophages, CD36 is thought to have a role as a scavenger receptor, mediating the phagocytosis of apoptotic cells and the endocytic uptake of oxidized low-density lipoproteins and fatty acids. The proposed topology of CD36 predicts that, of ten cysteine residues, six lie in the extracellular domain, whereas four are equally distributed in the two short terminal tails flanking the N-terminal and C-terminal hydrophobic stretches. Here we investigate the formation of intrachain disulphide bonds, on the basis of the assumption that the cysteine residues present in the luminal domains are generally oxidized, whereas those in the cytosol are reduced. As revealed by gel mobility-shift assays, disulphide bonds are present in the extracellular domain of the CD36 molecule. The formation of these bonds is required for the transport of CD36 from endoplasmic reticulum to Golgi. Furthermore reactive thiol groups are present in the CD36 sequence, which upon lysis form an intrachain extra loop as an artifact. This disulphide bond is not formed in either (1) truncated CD36 lacking the two C-terminal cysteine residues or (2) Triton X-100-insoluble wild-type CD36 molecules, suggesting that, in this fraction, the C-terminal thiol groups are modified.
Gruarin
P
P
DIBIT, San Raffaele Scientific Institute, via Olgettina 58, 20132 Milan, Italy.
Sitia
R
R
Alessio
M
M
D001692:Biological Transport; D018955:CD36 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D006801:Humans; D009000:Monocytes / Q000378:metabolism; D009154:Mutation; D010084:Oxidation-Reduction; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D017384:Sequence Deletion; D012694:Serine / Q000235:genetics; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism
D018955:CD36 Antigens; D004220:Disulfides; D011498:Protein Precursors; D012694:Serine; D003545:Cysteine