title: Proteomic profiling of the molecular targets of interactions of the mastoparan peptide Protopolybia MP-III at the level of endosomal membranes from rat mast cells.
abstract: It is well known that the activation of mast cells due to the binding of mastoparan to the G(α)  subunit of trimeric G proteins involves exocytosis regulation. However, experimental evidence in the literature indicates that mastoparan can also activate certain regulatory targets of exocytosis at the level of the mast cell endosomal membranes that have not yet been identified. Therefore, the aim of the present investigation was the proteomic identification of these targets. To achieve these objectives, mast cells were activated by the peptide Protopolybia MP-III, and the proteins of the endosomal membranes were converted to proteoliposomes using sonication. Proteins were separated from one another by affinity chromatography using proteoliposomes as analytes and Protopolybia MP III-immobilized Sepharose 4B resin as the ligand. This experimental approach, which used SDS-PAGE, in-gel trypsin digestion and proteomic analysis, permitted the identification of five endosomal proteins: Rho GTPase Cdc 42 and exocyst complex component 7 as components of the Ca(2+) -independent FcεRI-mediated exocytosis pathway, synaptosomal-associated protein 29, and GTP-binding protein Rab3D as components of the Ca(2+) -dependent FcεRI-mediated exocytosis pathway and Ras-related protein M-Ras, a protein that is related to the mediation of cell shaping and proliferation following exocytosis. The identification of the five proteins as targets of mastoparans may contribute in the near future to the use of this family of peptides as novel tools for dissecting the mechanism of exocytosis in mast cells.
authors: Center for the Study of Venoms and Venomous Animals, University of São Paulo State, Botucatu, SP, Brazil.
authors: Lucilene Delazari
authors: LD
authors: dos Santos
authors: José Roberto
authors: JR
authors: Aparecido dos Santos Pinto
authors: Anally Ribeiro da Silva
authors: AR
authors: Menegasso
authors: Daniel Menezes
authors: DM
authors: Saidemberg
authors: Ana Maria
authors: AM
authors: Caviquioli Garcia
authors: Mario Sergio
authors: MS
authors: Palma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015550:Cell Degranulation; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D011992:Endosomes / Q000201:enzymology / Q000378:metabolism*; D005089:Exocytosis; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D013058:Mass Spectrometry; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics; D051381:Rats; D014862:Wasp Venoms / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D014863:Wasps / Q000737:chemistry
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D014862:Wasp Venoms; C037500:mastoparan; D019204:GTP-Binding Proteins