title: CD36 is a ditopic glycoprotein with the N-terminal domain implicated in intracellular transport.
abstract: The CD36 receptor sequence predicts two hydrophobic domains located at the N- and C-termini of the protein, but there are conflicting reports as to whether the N-terminal uncleaved leader sequence functions as a transmembrane domain. To investigate the topology of CD36, we generated a panel of mutants lacking either one or both hydrophobic regions and analyzed their folding and transport in COS-7 cells. The N- and the C-terminal hydrophobic regions were both sufficient to anchor CD36 in the membrane, and a FLAG epitope inserted at the N-terminus was located intracellularly. These results indicate that CD36 adopts a ditopic configuration. Accordingly, neither N- nor C-terminal truncation mutants were secreted. Analysis with conformation-specific monoclonal antibodies showed that the N-terminal transmembrane domain truncated molecule was slowly transported through the exocytic pathway and largely accumulated intracellularly. Thus, dual membrane insertion dictates the correct topogenesis and seems to be necessary for efficient folding and intracellular transport.
authors.affiliation: DIBIT, San Raffaele Scientific Institute, via Olgettina 58, Milan, 20132, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Gruarin
authors.forename: R F
authors.initials: RF
authors.lastname: Thorne
authors.forename: D J
authors.initials: DJ
authors.lastname: Dorahy
authors.forename: G F
authors.initials: GF
authors.lastname: Burns
authors.forename: R
authors.initials: R
authors.lastname: Sitia
authors.forename: M
authors.initials: M
authors.lastname: Alessio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D018955:CD36 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D017931:DNA Primers; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017510:Protein Folding; D017386:Sequence Homology, Amino Acid
chemical_list: D018955:CD36 Antigens; D017931:DNA Primers