在毕赤酵母中表达的牛蜱抗原Bm95含有N-聚糖和O-聚糖的短链。

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

作者信息

González Luis J, Cremata José A, Guanche Yazmín, Ramos Yassel, Triguero Ada, Cabrera Gleysin, Montesino Raquel, Huerta Vivian, Pons Tirso, Boué Oscar, Farnós Omar, Rodríguez Manuel

机构信息

Division of Physical-Chemistry, Department of Proteomics, Havana, Cuba.

出版信息

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

DOI:10.1016/j.abb.2004.09.022

PMID:15542059

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.

摘要

Bm95是一种从微小牛蜱菌株中分离出的抗原，该菌株对用重组Bm86抗原（古巴HeberBiotec S.A.公司的Gavac）免疫的牛所产生的抗体敏感性较低。它是一种类似Bm86的表面蛋白，通过相似性分析发现其含有七个表皮生长因子（EGF）样结构域，并且在C末端区域有一个脂质结合糖基磷脂酰肌醇（GPI）锚定位点。在毕赤酵母中表达的重组（rBm95）蛋白的一级结构已通过液相色谱/质谱（LC/MS）完全验证。四个潜在的糖基化位点（天冬酰胺122、163、329和363）被短N聚糖部分糖基化，范围从Man(5)GlcNAc(2)到Man(9)GlcNAc(2)，其中Man(8 - 9)GlcNAc(2)最为丰富。O - 糖肽大多分布在蛋白质的N末端。第一个N - 糖基化位点（Asn(122)）位于EGF样结构域2和3之间，该区域也发现了O - 糖肽，另外两个N - 糖基化位点（Asn(329)和Asn(363)）位于EGF样结构域5和6之间，这一区域没有O - 糖基化的丝氨酸（Ser）或苏氨酸（Thr）。