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Serine scanning—A tool to prove the consequences of N-glycosylation of proteins

Paper ID Volume ID Publish Year Pages File Format Full-Text
25252 43563 2007 12 PDF Available
Title
Serine scanning—A tool to prove the consequences of N-glycosylation of proteins
Abstract

N-Glycosylation of proteins is a common posttranslational modification in eukaryotes. Often this results in enhanced protein stability through protection by the attached sugar moieties. Due to its 13 potential N-glycosylation motifs (N-X-T/S), recombinant hydroxynitrile lyase isoenzyme 5 from almonds (PaHNL5) is secreted by the heterologous host Pichia pastoris in a massively glycosylated form, and it shows extraordinary stability at low pH. The importance of N-glycosylation in general, and individual glycosylation sites in particular for stability at low pH were investigated. To identify especially important glycosylation sites asparagine from all N-X-S/T-motifs was replaced by serine. Thus, critical sites, which contributed to overall enzyme activity and/or stability, were identified individually. One glycosylation site revealed to be essential for stability at low pH. After enzymatic deglycosylation, leaving only one acetylglucosamine attached to asparagines, PaHNL5 retained most of its stability at low pH. Protonation effects in the active site as well as higher-order aggregational events upon incubation in low pH were excluded. This study provides evidence for the interconnection of N-glycosylation and stability at low pH for PaHNL5. Moreover, serine scanning was proven to be applicable for quick identification of critical glycosylation sites.

Keywords
pH stability; N-Glycosylation; Serine scanning; Pichia pastoris; Hydroxynitrile lyase (HNL); Tunicamycin
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Serine scanning—A tool to prove the consequences of N-glycosylation of proteins
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 129, Issue 1, 30 March 2007, Pages 50–61
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us