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Resources and Methods for Engineering "Designer" Glycan-Binding Proteins.

Author(s): Warkentin R, Kwan DH

This review provides information on available methods for engineering glycan-binding proteins (GBP). Glycans are involved in a variety of physiological functions and are found in all domains of life and viruses. Due to their wide range of functions, GBPs ha...

Article GUID: 33450899
Enzymatic Synthesis of a Fluorogenic Reporter Substrate and the Development of a High-Throughput Assay for Fucosyltransferase VIII Provide a Toolkit to Probe and Inhibit Core Fucosylation.

Author(s): Soroko M, Kwan DH

Biochemistry. 2020 Jun 01;: Authors: Soroko M, Kwan DH

Article GUID: 32441090
Structure-Guided Directed Evolution of Glycosidases: A Case Study in Engineering a Blood Group Antigen-Cleaving Enzyme.

Author(s): Kwan DH

Methods Enzymol. 2017;597:25-53 Authors: Kwan DH

Article GUID: 28935105
Title:Structure-Guided Directed Evolution of Glycosidases: A Case Study in Engineering a Blood Group Antigen-Cleaving Enzyme.
Authors:Kwan DH
Link:https://www.ncbi.nlm.nih.gov/pubmed/28935105?dopt=Abstract
DOI:10.1016/bs.mie.2017.06.002
Category:Methods Enzymol
PMID:28935105
Dept Affiliation: GENOMICS
1 Centre for Applied Synthetic Biology, Centre for Structural and Functional Genomics, Concordia University, Montréal, Québec, Canada. Electronic address: david.kwan@concordia.ca.

Description:

Structure-Guided Directed Evolution of Glycosidases: A Case Study in Engineering a Blood Group Antigen-Cleaving Enzyme.

Methods Enzymol. 2017;597:25-53

Authors: Kwan DH

Abstract

Directed evolution is an incredibly powerful strategy for engineering enzyme function. Applying this approach to glycosidases offers enormous potential for the development of highly specialized tools in chemical glycobiology. Performing enzyme directed evolution requires the generation, by random mutagenesis, of mutant libraries from which large numbers of variant enzymes must be screened in high-throughput assays. A structure-guided "semirational" method for library creation allows researchers to target specific amino acid positions for mutagenesis, concentrating mutations where they might be most effective in order to produce mutant libraries of a manageable size, minimizing screening effort while maximizing the chances of finding improved mutants. Well-designed assays, which may use specially prepared substrates, enable efficient screening of these mutant libraries. This chapter will detail general methods in the structure-guided directed evolution of glycosidases, which have previously been employed in engineering a blood group antigen-cleaving enzyme.

PMID: 28935105 [PubMed - indexed for MEDLINE]