title: Use of proteomic methodology for the characterization of human milk fat globular membrane proteins.
abstract: Characterization of the major human milk fat globular membrane proteins was carried out using proteomic techniques comprising two-dimensional polyacrylamide gel electrophoresis, followed by in situ PNGase F and trypsin digestion. Matrix-assisted laser desorption/ionization quadrupole time-of-flight and electrospray ionization mass spectrometry identified seven major protein components: alpha-lactalbumin, lysozyme precursor, beta-casein, clusterin, lactotransferrin, polymeric immunoglobulin receptor precursor, and human milk fat globule EGF-factor 8 protein. Sequence information on the protein-associated glycans was determined by matrix-assisted laser desorption-ionization quadrupole time-of-flight hybrid mass spectrometry. This glycan analysis revealed interesting fucosylation branching patterns which may be influential in maternal protection of the newborn against bacterial and viral pathogenic attack.
authors.affiliation: New Frontiers Science Park, GlaxoSmithKline, Third Avenue, Harlow, Essex, CM19 5AW, United Kingdom.
authors.forename: Joanne
authors.initials: J
authors.lastname: Charlwood
authors.forename: Sarah
authors.initials: S
authors.lastname: Hanrahan
authors.forename: Richard
authors.initials: R
authors.lastname: Tyldesley
authors.forename: James
authors.initials: J
authors.lastname: Langridge
authors.forename: Miriam
authors.initials: M
authors.lastname: Dwek
authors.forename: Patrick
authors.initials: P
authors.lastname: Camilleri
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D002364:Caseins / Q000032:analysis; D051152:Clusterin; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004792:Enzyme Precursors; D004815:Epidermal Growth Factor / Q000032:analysis; D005260:Female; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007768:Lactalbumin / Q000032:analysis; D007781:Lactoferrin / Q000032:analysis; D008565:Membrane Proteins / Q000737:chemistry; D008894:Milk Proteins / Q000032:analysis*; D008895:Milk, Human / Q000737:chemistry*; D018832:Molecular Chaperones / Q000032:analysis; D008969:Molecular Sequence Data; D009113:Muramidase / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis* / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis*; D019056:Receptors, Polymeric Immunoglobulin / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: C495242:CLU protein, human; D002364:Caseins; D051152:Clusterin; D004792:Enzyme Precursors; D006023:Glycoproteins; D008565:Membrane Proteins; D008894:Milk Proteins; D018832:Molecular Chaperones; D011134:Polysaccharides; D020543:Proteome; D019056:Receptors, Polymeric Immunoglobulin; D004815:Epidermal Growth Factor; D007768:Lactalbumin; D009113:Muramidase; D007781:Lactoferrin