Site-specific glycosylation of bovine butyrophilin.

Site-specific glycosylation of bovine butyrophilin.
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牛嗜丁蛋白的位点特异性糖基化。

DOI:
10.1093/oxfordjournals.jbchem.a124702
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发表时间:
1995
影响因子:
2.7
通讯作者:
Furukawa,K
Furukawa,K
中科院分区:
生物学4区
文献类型:
--
作者:
Sato,T;Takio,K;Kobata,A;Greenwalt,DE;Furukawa,K

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我们以前的研究表明,来自乳脂球膜(MFGM)的大多数牛糖蛋白的N-连接糖链含有GalNAcβ1→4GlcNAc基团[Satoet al.(1993)J.Biochem.114,890-900]。由于二糖结构的表达受糖基化位点附近的肽序列的影响[Smith和Baenziger(1992)Proc.Natl. Acad. Sci. USA 89,329-333],使用牛亲酪蛋白(一种具有已知一级结构的主要MFGM糖蛋白)研究N-乙酰半乳糖胺化糖链的位点特异性。用无色杆菌蛋白酶Ⅰ酶解该蛋白,得到两个糖肽片段,它们含有连接在Asn-55或Asn-215残基上的N-连接糖链。通过肼解从每个糖肽释放的糖链用NaB 3 H4还原。通过连续外切糖苷酶消化和甲基化分析对寡糖进行结构分析,结果表明,Asn-55连接寡糖中仅包含具有GalNAcβl→4GlcNAc结构的复合型糖链,而Asn-215连接寡糖中仅包含先前在牛MFGM糖蛋白中检测到的新型杂交型糖链。结果表明,butyrophilin的糖基化以位点特异性方式发生。
Our previous studies showed that theN-linked sugar chains of most bovine glycoproteins from milk fat globule membranes (MFGM) contain the GalNAcβ1→4GlcNAc group [Satoet al. (1993)J. Biochem. 114, 890–900]. Since expression of the disaccharide structure is influenced by peptide sequences near the glycosylation sites [Smith and Baenziger (1992)Proc. Natl. Acad. Sci. USA 89, 329–333], the site-specificity of theN-acetylgalactosaminylated sugar chains was investigated using bovine butyrophilin, a major MFGM glycoprotein with known primary structure. Two glycopeptide fragments which contained theN-linked sugar chains linked to either Asn-55 or Asn-215 residue were obtained by digestion of the protein withAchromobacterprotease I. The sugar chains released from each glycopeptide by hydrazinolysis were reduced with NaB3H4. Structural analyses of the oligosaccharides by sequential exoglycosidase digestion and methylation analysis revealed that only complex-type sugar chains with the GalNAcβl→4GlcNAc structure are included in Asn-55-linked oligosaccharides, while only novel hybrid-type sugar chains detected previously in bovine MFGM glycoproteins are included in Asn-215-linked oligosaccharides. The results show that the glycosylation of butyrophilin occurs in a site-specific manner.
DOI: 10.1111/j.1432-1033.1992.tb17362.x
发表时间: 1992-11-01
期刊: EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子: --
作者:
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