Restoration of lectin activity to a non‐glycosylated ricin B chain mutant by the introduction of a novel N‐glycosylation site

FEBS Letters

Chaddock, John A.¹  Zhan, Jinbiao¹  Roberts, Lynne M.¹  Lord, J.Michael¹  de Sousa, Michelle¹ 

[1] Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK

关键词: Ricin B chain;    Lectin activity;    N-glycosylation;   

DOI  :  10.1016/S0014-5793(97)00341-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Ricin B chain (RTB) is an N-glycosylated, galactose-specific lectin. Removal of the two native N-glycosylation sites at Asn⁹⁵ and Asn¹³⁵ by site-directed mutagenesis generated a recombinant protein devoid of lectin activity. Two novel N-glycosylation sites were introduced into RTB at Asn⁴² and Asn¹²³, either singly or in combination. Microinjection of pre-RTB transcripts into Xenopus oocytes showed that these novel sites became glycosylated in vivo. The single oligosaccharide site chain at Asn⁴² restored lectin activity to RTB, whereas glycosylation at Asn¹²³ or simultaneous glycosylation at Asn⁴² and Asn¹²³ failed to do so.

【 授权许可】

Unknown