α2-6-Neosialidase: A Sialyltransferase Mutant as a Sialyl Linkage-Specific Sialidase

John B. McArthur, Hai Yu, Nova Tasnima, Christie M. Lee, Andrew J Fisher, Xi Chen

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The lack of α2-6-linkage specific sialidases limits the structural and functional studies of sialic-acid-containing molecules. Photobacterium damselae α2-6-sialyltransferase (Pd2,6ST) was shown previously to have α2-6-specific, but weak, sialidase activity. Here, we develop a high-throughput blue-white colony screening method to identify Pd2,6ST mutants with improved α2-6-sialidase activity from mutant libraries generated by sequential saturation mutagenesis. A triple mutant (Pd2,6ST S232L/T356S/W361F) has been identified with 100-fold improved activity, high α2-6-sialyl linkage selectivity, and ability to cleave two common sialic acid forms, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). It is a valuable tool for sialoglycan structural analysis and functional characterization. The sequential saturation mutagenesis and screening strategy developed here can be explored to evolve other linkage-specific neoglycosidases from the corresponding glycosyltransferases.

Original languageEnglish (US)
Pages (from-to)1228-1234
Number of pages7
JournalACS Chemical Biology
Volume13
Issue number5
DOIs
StatePublished - May 18 2018

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Photobacterium
Sialyltransferases
Neuraminidase
N-Acetylneuraminic Acid
Mutagenesis
Screening
Glycosyltransferases
Structural analysis
Throughput
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

α2-6-Neosialidase : A Sialyltransferase Mutant as a Sialyl Linkage-Specific Sialidase. / McArthur, John B.; Yu, Hai; Tasnima, Nova; Lee, Christie M.; Fisher, Andrew J; Chen, Xi.

In: ACS Chemical Biology, Vol. 13, No. 5, 18.05.2018, p. 1228-1234.

Research output: Contribution to journalArticle

McArthur, John B. ; Yu, Hai ; Tasnima, Nova ; Lee, Christie M. ; Fisher, Andrew J ; Chen, Xi. / α2-6-Neosialidase : A Sialyltransferase Mutant as a Sialyl Linkage-Specific Sialidase. In: ACS Chemical Biology. 2018 ; Vol. 13, No. 5. pp. 1228-1234.
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