Use of proteomic methodology for the characterization of human milk fat globular membrane proteins.
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.
Charlwood
Joanne
J
New Frontiers Science Park, GlaxoSmithKline, Third Avenue, Harlow, Essex, CM19 5AW, United Kingdom.
Hanrahan
Sarah
S
Tyldesley
Richard
R
Langridge
James
J
Dwek
Miriam
M
Camilleri
Patrick
P
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*
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