The cattle tick antigen, Bm95, expressed in Pichia pastoris contains short chains of N- and O-glycans

Arch Biochem Biophys. 2004 Dec 15;432(2):205-11. doi: 10.1016/j.abb.2004.09.022.

Abstract

Bm95 is an antigen isolated from Boophilus microplus strains with low susceptibility to antibodies developed in cattle vaccinated with the recombinant Bm86 antigen (Gavac, HeberBiotec S.A., Cuba). It is a Bm86-like surface protein, which by similarity contains seven EGF-like domains and a lipid-binding GPI-anchor site at the C-terminal region. The primary structure of the recombinant (rBm95) protein expressed in Pichia pastoris was completely verified by LC/MS. The four potential glycosylation sites (Asn 122, 163, 329, and 363) are glycosylated partially with short N-glycans, from Man(5)GlcNAc(2) to Man(9)GlcNAc(2) of which, Man(8-9)GlcNAc(2) were the most abundant. O-Glycopeptides are distributed mostly towards the protein N-terminus. While the first N-glycosylated site (Asn(122)) is located between EGF-like domains 2 and 3, where the O-glycopeptides were found, two other N-glycosylated sites (Asn(329) and Asn(363)) are located between EGF-like domains 5 and 6, a region devoid of O-glycosylated Ser or Thr.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cloning, Molecular
  • Glycosylation
  • Membrane Glycoproteins / chemistry*
  • Membrane Glycoproteins / genetics*
  • Molecular Sequence Data
  • Pichia / genetics
  • Pichia / metabolism
  • Polysaccharides / chemistry*
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics*
  • Structure-Activity Relationship
  • Ticks / genetics
  • Ticks / metabolism*
  • Vaccines / chemistry*
  • Vaccines / genetics*

Substances

  • BM86 protein, Boophilus
  • Membrane Glycoproteins
  • Polysaccharides
  • Recombinant Proteins
  • Vaccines

Associated data

  • SWISSPROT/P20736