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

Eriksson, Torny LU ; Stals, Ingeborg ; Collén, Anna LU ; Tjerneld, Folke LU ; Claeyssens, Marc ; Stålbrand, Henrik LU and Brumer, Harry (2004) In European Journal of Biochemistry 271(7). p.1266-1276
Abstract
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. GlcNAc2Man5 was identified as the predominant N-glycan, although lesser amounts of GlcNAc2Man7 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... (More)
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. GlcNAc2Man5 was identified as the predominant N-glycan, although lesser amounts of GlcNAc2Man7 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. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
N-glycan, protein glycosylation, O-glycan, Tricho-derma reesei, cellulase.
in
European Journal of Biochemistry
volume
271
issue
7
pages
1266 - 1276
publisher
Wiley-Blackwell
external identifiers
  • pmid:15030476
  • wos:000220355100004
  • scopus:1942455827
  • pmid:15030476
ISSN
0014-2956
DOI
10.1111/j.1432-1033.2004.04031.x
language
English
LU publication?
yes
id
1ebce2c9-67cc-4ebd-a4dc-4450bf3c1dd8 (old id 121112)
date added to LUP
2016-04-01 15:53:17
date last changed
2022-04-15 00:39:31
@article{1ebce2c9-67cc-4ebd-a4dc-4450bf3c1dd8,
  abstract     = {{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. GlcNAc2Man5 was identified as the predominant N-glycan, although lesser amounts of GlcNAc2Man7 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.}},
  author       = {{Eriksson, Torny and Stals, Ingeborg and Collén, Anna and Tjerneld, Folke and Claeyssens, Marc and Stålbrand, Henrik and Brumer, Harry}},
  issn         = {{0014-2956}},
  keywords     = {{N-glycan; protein glycosylation; O-glycan; Tricho-derma reesei; cellulase.}},
  language     = {{eng}},
  number       = {{7}},
  pages        = {{1266--1276}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{European Journal of Biochemistry}},
  title        = {{Heterogeneity of homologously expressed Hypocrea jecorina (Trichoderma reesei) Cel7B catalytic module.}},
  url          = {{http://dx.doi.org/10.1111/j.1432-1033.2004.04031.x}},
  doi          = {{10.1111/j.1432-1033.2004.04031.x}},
  volume       = {{271}},
  year         = {{2004}},
}