The cattle tick antigen, Bm95, expressed in Pichia pastoris contains short chains of N- and O-glycans.
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.
González
Luis J
LJ
Division of Physical-Chemistry, Department of Proteomics, Havana, Cuba.
Cremata
José A
JA
Guanche
Yazmín
Y
Ramos
Yassel
Y
Triguero
Ada
A
Cabrera
Gleysin
G
Montesino
Raquel
R
Huerta
Vivian
V
Pons
Tirso
T
Boué
Oscar
O
Farnós
Omar
O
Rodríguez
Manuel
M
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics*; D013329:Structure-Activity Relationship; D013987:Ticks / Q000235:genetics / Q000378:metabolism*; D014612:Vaccines / Q000737:chemistry* / Q000235:genetics*
C085065:BM86 protein, Boophilus; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D014612:Vaccines