Heterogeneity of homologously expressed Hypocrea jecorina (Trichoderma reesei) Cel7B catalytic module

被引：25

作者：

Eriksson, T

Stals, I

Collén, A

Tjerneld, F

Claeyssens, M

Stålbrand, H

Brumer, H ^{[1
]}

机构：

[1] Royal Inst Technol, Dept Biotechnol, AlbaNova Univ Ctr, S-10691 Stockholm, Sweden

[2] Lund Univ, Ctr Chem & Chem Engn, Dept Biochem, S-22100 Lund, Sweden

[3] Univ Ghent, Dept Biochem Physiol & Microbiol, Biochem Lab, Ghent, Belgium

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 2004年 / 271卷 / 07期

关键词：

protein glycosylation; O-glycan; N-glycan; Trichoderma reesei; cellulase;

D O I：

10.1111/j.1432-1033.2004.04031.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The catalytic module of Hypocrea jecorina (previously Trichoderma reesei) Cel7B was homologously expressed by transformation of strain QM9414. Post-translational modifications in purified Cel7B preparations were analysed by enzymatic digestions, high performance chromatography, mass spectrometry and site-directed mutagenesis. Of the five potential sites found in the wild-type enzyme, only Asn56 and Asn182 were found to be N-glycosylated. GlcNAc(2)Man(5) was identified as the predominant N-glycan, although lesser amounts of GlcNAc(2)Man(7) and glycans carrying a mannophosphodiester bond were also detected. Repartition of neutral and charged glycan structures over the two glycosylation sites mainly accounts for the observed microheterogeneity of the protein. However, partial deamidation of Asn259 and a partially occupied O-glycosylation site give rise to further complexity in enzyme preparations.

引用

页码：1266 / 1276

页数：11

共 38 条

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