title: Characterization of β-lactoglobulin from buffalo (Bubalus bubalis) colostrum and its possible interaction with erythrocyte lipocalin-interacting membrane receptor.
abstract: Lipocalins form a widespread class of proteins involved in the transport of weakly soluble vitamins, hormones or hydrophobic molecules. β-lactoglobulin (BLG-col), a major lipocalin present in whey was purified and characterized from buffalo colostrum. The molecular weight of BLG-col as determined by Liquid chromatography -electrospray ionization mass spectrometry (LC-ESI-MS) was 18.257 kDa and the peptide mass fingerprint of the purified protein revealed 67% sequence homology to buffalo milk β-lg. The N-terminal-IIVTQ and LC-ESI-collision-induced dissociation-Electron transfer dissociation mass spectrometry/mass spectrometry analyses of doubly (m/z 1156(+2)) and triply (m/z 546(+3)) charged ion pairs corresponding to VYVEELKPTPEGDLEILLQK (41-60) and TPEVDDEALEKFDK (125-138) sequences confirmed the identity of BLG-col. Using these peptide sequences, the location of a gene encoding for BLG-col was identified on chromosome 11 at 11q28 loci of bovine genome. The unique property of the BLG-col isolated from buffalo colostrum was its strong and specific haemagglutinating activity with 'O' blood of human erythrocytes with 10,309 HAU/mg protein. The cell surface localization of BLG-col on human erythrocytes was confirmed by immunocytochemistry and the specificity of interaction was established by immunoblot analysis of human erythrocyte membrane proteins. Based on these observations, we suggest the presence of lipocalin receptor (70 kDa) on human erythrocyte membrane and the multiple sequence alignment supported structural diversity among lipocalin receptors.
authors.affiliation: Department of Studies in Biotechnology, University of Mysore, Manasagangotri, Mysore 570 006, Karnataka, India.
authors.forename: Rohit A
authors.initials: RA
authors.lastname: Chougule
authors.forename: Huligerepura S
authors.initials: HS
authors.lastname: Aparna
mesh_terms: D000017:ABO Blood-Group System / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D002020:Buffaloes / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D003126:Colostrum / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006388:Hemagglutinins / Q000276:immunology; D006801:Humans; D007782:Lactoglobulins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D054834:Lipocalins / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011247:Pregnancy; D011971:Receptors, Immunologic / Q000276:immunology*; D063267:Sialic Acid Binding Ig-like Lectin 1
chemical_list: D000017:ABO Blood-Group System; D006388:Hemagglutinins; D007782:Lactoglobulins; D054834:Lipocalins; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; C571166:SIGLEC1 protein, human; D063267:Sialic Acid Binding Ig-like Lectin 1