A short peptide motif at the carboxyl terminus is required for incorporation of the integral membrane MAL protein to glycolipid-enriched membranes.
The MAL (VIP17, MVP17) proteolipid, an integral membrane protein with specific residence in glycolipid-enriched membrane (GEM) microdomains, has been recently proposed as a component of the protein machinery for GEM vesiculation. In this work, we have searched the COOH terminus of MAL for sorting determinants responsible for targeting to GEMs. This has allowed the identification of the sequence Leu-Ile-Arg-Trp (LIRW) as necessary for the access of MAL to GEMs. This motif requires at least one additional amino acid at its COOH end for full effectiveness. The arginine within the LIRW motif is the most crucial residue for targeting to GEMs, tryptophan replacement affects targeting to a lesser extent, and the leucine-isoleucine pair tolerates substitution by valine, but not by alanine, without effect on targeting. Pulse-chase experiments indicate that the LIRW tetrapeptide is required for access to GEMs early after MAL biosynthesis. Interestingly, the loss of the capacity of the MAL protein to be incorporated into GEMs correlated with the loss of its response to brefeldin A treatment. This is the first identification of a juxtamembrane peptide motif required for incorporation of an integral membrane protein into GEMs.
Puertollano
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R
Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, Cantoblanco, 28049-Madrid, Spain.
Alonso
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002460:Cell Line; D017931:DNA Primers; D006017:Glycolipids / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D026901:Membrane Transport Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009185:Myelin Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D011485:Protein Binding; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism*; D017384:Sequence Deletion
D017931:DNA Primers; D006017:Glycolipids; C569026:MAL protein, human; C569027:Mal protein, mouse; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D009185:Myelin Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D011510:Proteolipids