Presence of a glycan at a potential N-glycosylation site, Asn-281, of bovine lactoferrin.
This work was performed to clarify the differences in glycan moieties between multiple molecular mass forms of bovine lactoferrins (bovine lactoferrins-a and -b). After digestion of both bovine lactoferrins with cyanogen bromide and V8 protease, glycopeptides were successively purified by concavalin A affinity chromatography and HPLC on an octadecylsilyl column. Four glycopeptides glycosylated at Asn-233, -368, -476, and -545 were obtained from both hydrolysates of bovine lactoferrins-a and -b. On the other hand, a glycopeptide glycosylated at Asn-281 was only detected in hydrolysate of bovine lactoferrin-a, indicating that bovine lactoferrin-a possessed five N-glycosylated sites. The glycan linked to Asn-281 of bovine lactoferrin-a was found to consist of fucose, galactose, and N-acetylgalactosamine in addition to mannose and N-acetylglucosamine. HPLC analysis of this glycan on a normal phase column showed that peaks of several glycans were detected. These glycans changed to one major glycan consisting of only mannose and N-acetylglucosamine on exoglycosidase digestion. From these results, this glycan seemed to be of the complex type and possess heterogeneous structure.
Wei
Z
Z
Faculty of Applied Biological Science, Hiroshima University, Japan.
Nishimura
T
T
Yoshida
S
S
D000116:Acetylgalactosamine / Q000032:analysis; D000117:Acetylglucosamine / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine / Q000378:metabolism*; D001665:Binding Sites; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002845:Chromatography / Q000379:methods; D003488:Cyanogen Bromide; D005643:Fucose / Q000032:analysis; D005690:Galactose / Q000032:analysis; D006031:Glycosylation; D007781:Lactoferrin / Q000032:analysis* / Q000737:chemistry; D008358:Mannose / Q000032:analysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D012697:Serine Endopeptidases / Q000378:metabolism
D010446:Peptide Fragments; D011134:Polysaccharides; D005643:Fucose; D001216:Asparagine; D007781:Lactoferrin; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D000116:Acetylgalactosamine; D003488:Cyanogen Bromide; D008358:Mannose; D000117:Acetylglucosamine; D005690:Galactose