Glycan specificity of neuraminidases determined in microarray format

Janet E. McCombs, Jason P. Diaz, Kevin J. Luebke, Jennifer J. Kohler

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Neuraminidases hydrolytically remove sialic acids from glycoconjugates. Neuraminidases are produced by both humans and their pathogens, and function in normal physiology and in pathological events. Identification of neuraminidase substrates is needed to reveal their mechanism of action, but high-throughput methods to determine glycan specificity of neuraminidases are limited. Here we use two glycan labeling reactions to monitor neuraminidase activity toward glycan substrates. While both periodate oxidation and aniline-catalyzed oxime ligation (PAL) and galactose oxidase and aniline-catalyzed oxime ligation (GAL) can be used to monitor neuraminidase activity toward glycans in microtiter plates, only GAL accurately measured neuraminidase activity toward glycans displayed on a commercial glass slide microarray. Using GAL, we confirm known linkage specificities of three pneumococcal neuraminidases and obtain new information about underlying glycan specificity.

Original languageEnglish (US)
Pages (from-to)31-40
Number of pages10
JournalCarbohydrate Research
Volume428
DOIs
StatePublished - Jun 16 2016

Fingerprint

Neuraminidase
Microarrays
Polysaccharides
Oximes
Ligation
Galactose Oxidase
Sialic Acids
Glycoconjugates
Physiology
Pathogens
Substrates
Labeling
Glass
Throughput
Oxidation

Keywords

  • Chemoenzymatic labeling
  • Microarray
  • Neuraminidase
  • Sialic acids

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Organic Chemistry

Cite this

Glycan specificity of neuraminidases determined in microarray format. / McCombs, Janet E.; Diaz, Jason P.; Luebke, Kevin J.; Kohler, Jennifer J.

In: Carbohydrate Research, Vol. 428, 16.06.2016, p. 31-40.

Research output: Contribution to journalArticle

McCombs, Janet E. ; Diaz, Jason P. ; Luebke, Kevin J. ; Kohler, Jennifer J. / Glycan specificity of neuraminidases determined in microarray format. In: Carbohydrate Research. 2016 ; Vol. 428. pp. 31-40.
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