A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses

Xuezheng Song, Hai Yu, Xi Chen, Yi Lasanajak, Mary M. Tappert, Gillian M. Air, Vinod K. Tiwari, Hongzhi Cao, Harshal A. Chokhawala, Haojie Zheng, Richard D. Cummings, David F. Smith

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

Many glycan-binding proteins in animals and pathogens recognize sialic acid or its modified forms, but their molecular recognition is poorly understood. Here we describe studies on sialic acid recognition using a novel sialylated glycan microarray containing modified sialic acids presented on different glycan backbones. Glycans terminating in β-linked galactose at the nonreducing end and with an alkylamine-containing fluorophore at the reducing end were sialylated by a one-pot three-enzyme system to generate α2-3- and α2-6-linked sialyl glycans with 16 modified sialic acids. The resulting 77 sialyl glycans were purified and quantified, characterized by mass spectrometry, covalently printed on activated slides, and interrogated with a number of key sialic acid-binding proteins and viruses. Sialic acid recognition by the sialic acid-binding lectins Sambucus nigra agglutinin and Maackia amurensis lectin-I, which are routinely used for detecting α2-6- and α2-3-linked sialic acids, are affected by sialic acid modifications, and both lectins bind glycans terminating with 2-keto-3-deoxy-D-glycero-D- galactonononic acid (Kdn) and Kdn derivatives stronger than the derivatives of morecommonN-acetylneuraminic acid (Neu5Ac) andN-glycolylneuraminic acid (Neu5Gc). Three human parainfluenza viruses bind to glycans terminating with Neu5Ac or Neu5Gc and some of their derivatives but not to Kdn and its derivatives. Influenza A virus also does not bind glycans terminating in Kdn or Kdn derivatives. An especially novel aspect of human influenza A virus binding is its ability to equivalently recognize glycans terminated with either α2-6-linked Neu5Ac9Lt or α2-6-linked Neu5Ac. Our results demonstrate the utility of this sialylated glycan microarray to investigate the biological importance of modified sialic acids in protein-glycan interactions.

Original languageEnglish (US)
Pages (from-to)31610-31622
Number of pages13
JournalJournal of Biological Chemistry
Volume286
Issue number36
DOIs
StatePublished - Sep 9 2011

Fingerprint

Sialic Acids
Microarrays
Viruses
Polysaccharides
N-Acetylneuraminic Acid
Proteins
Derivatives
Acids
Influenza A virus
Lectins
Carrier Proteins
Maackia
Virus Attachment
Paramyxoviridae Infections
Molecular recognition
Fluorophores
Agglutinins
Pathogens
Galactose
Human Influenza

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses. / Song, Xuezheng; Yu, Hai; Chen, Xi; Lasanajak, Yi; Tappert, Mary M.; Air, Gillian M.; Tiwari, Vinod K.; Cao, Hongzhi; Chokhawala, Harshal A.; Zheng, Haojie; Cummings, Richard D.; Smith, David F.

In: Journal of Biological Chemistry, Vol. 286, No. 36, 09.09.2011, p. 31610-31622.

Research output: Contribution to journalArticle

Song, X, Yu, H, Chen, X, Lasanajak, Y, Tappert, MM, Air, GM, Tiwari, VK, Cao, H, Chokhawala, HA, Zheng, H, Cummings, RD & Smith, DF 2011, 'A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses', Journal of Biological Chemistry, vol. 286, no. 36, pp. 31610-31622. https://doi.org/10.1074/jbc.M111.274217
Song, Xuezheng ; Yu, Hai ; Chen, Xi ; Lasanajak, Yi ; Tappert, Mary M. ; Air, Gillian M. ; Tiwari, Vinod K. ; Cao, Hongzhi ; Chokhawala, Harshal A. ; Zheng, Haojie ; Cummings, Richard D. ; Smith, David F. / A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 36. pp. 31610-31622.
@article{d5c3c18082ad42b788f95c2a7a2e1546,
title = "A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses",
abstract = "Many glycan-binding proteins in animals and pathogens recognize sialic acid or its modified forms, but their molecular recognition is poorly understood. Here we describe studies on sialic acid recognition using a novel sialylated glycan microarray containing modified sialic acids presented on different glycan backbones. Glycans terminating in β-linked galactose at the nonreducing end and with an alkylamine-containing fluorophore at the reducing end were sialylated by a one-pot three-enzyme system to generate α2-3- and α2-6-linked sialyl glycans with 16 modified sialic acids. The resulting 77 sialyl glycans were purified and quantified, characterized by mass spectrometry, covalently printed on activated slides, and interrogated with a number of key sialic acid-binding proteins and viruses. Sialic acid recognition by the sialic acid-binding lectins Sambucus nigra agglutinin and Maackia amurensis lectin-I, which are routinely used for detecting α2-6- and α2-3-linked sialic acids, are affected by sialic acid modifications, and both lectins bind glycans terminating with 2-keto-3-deoxy-D-glycero-D- galactonononic acid (Kdn) and Kdn derivatives stronger than the derivatives of morecommonN-acetylneuraminic acid (Neu5Ac) andN-glycolylneuraminic acid (Neu5Gc). Three human parainfluenza viruses bind to glycans terminating with Neu5Ac or Neu5Gc and some of their derivatives but not to Kdn and its derivatives. Influenza A virus also does not bind glycans terminating in Kdn or Kdn derivatives. An especially novel aspect of human influenza A virus binding is its ability to equivalently recognize glycans terminated with either α2-6-linked Neu5Ac9Lt or α2-6-linked Neu5Ac. Our results demonstrate the utility of this sialylated glycan microarray to investigate the biological importance of modified sialic acids in protein-glycan interactions.",
author = "Xuezheng Song and Hai Yu and Xi Chen and Yi Lasanajak and Tappert, {Mary M.} and Air, {Gillian M.} and Tiwari, {Vinod K.} and Hongzhi Cao and Chokhawala, {Harshal A.} and Haojie Zheng and Cummings, {Richard D.} and Smith, {David F.}",
year = "2011",
month = "9",
day = "9",
doi = "10.1074/jbc.M111.274217",
language = "English (US)",
volume = "286",
pages = "31610--31622",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "36",

}

TY - JOUR

T1 - A sialylated glycan microarray reveals novel interactions of modified sialic acids with proteins and viruses

AU - Song, Xuezheng

AU - Yu, Hai

AU - Chen, Xi

AU - Lasanajak, Yi

AU - Tappert, Mary M.

AU - Air, Gillian M.

AU - Tiwari, Vinod K.

AU - Cao, Hongzhi

AU - Chokhawala, Harshal A.

AU - Zheng, Haojie

AU - Cummings, Richard D.

AU - Smith, David F.

PY - 2011/9/9

Y1 - 2011/9/9

N2 - Many glycan-binding proteins in animals and pathogens recognize sialic acid or its modified forms, but their molecular recognition is poorly understood. Here we describe studies on sialic acid recognition using a novel sialylated glycan microarray containing modified sialic acids presented on different glycan backbones. Glycans terminating in β-linked galactose at the nonreducing end and with an alkylamine-containing fluorophore at the reducing end were sialylated by a one-pot three-enzyme system to generate α2-3- and α2-6-linked sialyl glycans with 16 modified sialic acids. The resulting 77 sialyl glycans were purified and quantified, characterized by mass spectrometry, covalently printed on activated slides, and interrogated with a number of key sialic acid-binding proteins and viruses. Sialic acid recognition by the sialic acid-binding lectins Sambucus nigra agglutinin and Maackia amurensis lectin-I, which are routinely used for detecting α2-6- and α2-3-linked sialic acids, are affected by sialic acid modifications, and both lectins bind glycans terminating with 2-keto-3-deoxy-D-glycero-D- galactonononic acid (Kdn) and Kdn derivatives stronger than the derivatives of morecommonN-acetylneuraminic acid (Neu5Ac) andN-glycolylneuraminic acid (Neu5Gc). Three human parainfluenza viruses bind to glycans terminating with Neu5Ac or Neu5Gc and some of their derivatives but not to Kdn and its derivatives. Influenza A virus also does not bind glycans terminating in Kdn or Kdn derivatives. An especially novel aspect of human influenza A virus binding is its ability to equivalently recognize glycans terminated with either α2-6-linked Neu5Ac9Lt or α2-6-linked Neu5Ac. Our results demonstrate the utility of this sialylated glycan microarray to investigate the biological importance of modified sialic acids in protein-glycan interactions.

AB - Many glycan-binding proteins in animals and pathogens recognize sialic acid or its modified forms, but their molecular recognition is poorly understood. Here we describe studies on sialic acid recognition using a novel sialylated glycan microarray containing modified sialic acids presented on different glycan backbones. Glycans terminating in β-linked galactose at the nonreducing end and with an alkylamine-containing fluorophore at the reducing end were sialylated by a one-pot three-enzyme system to generate α2-3- and α2-6-linked sialyl glycans with 16 modified sialic acids. The resulting 77 sialyl glycans were purified and quantified, characterized by mass spectrometry, covalently printed on activated slides, and interrogated with a number of key sialic acid-binding proteins and viruses. Sialic acid recognition by the sialic acid-binding lectins Sambucus nigra agglutinin and Maackia amurensis lectin-I, which are routinely used for detecting α2-6- and α2-3-linked sialic acids, are affected by sialic acid modifications, and both lectins bind glycans terminating with 2-keto-3-deoxy-D-glycero-D- galactonononic acid (Kdn) and Kdn derivatives stronger than the derivatives of morecommonN-acetylneuraminic acid (Neu5Ac) andN-glycolylneuraminic acid (Neu5Gc). Three human parainfluenza viruses bind to glycans terminating with Neu5Ac or Neu5Gc and some of their derivatives but not to Kdn and its derivatives. Influenza A virus also does not bind glycans terminating in Kdn or Kdn derivatives. An especially novel aspect of human influenza A virus binding is its ability to equivalently recognize glycans terminated with either α2-6-linked Neu5Ac9Lt or α2-6-linked Neu5Ac. Our results demonstrate the utility of this sialylated glycan microarray to investigate the biological importance of modified sialic acids in protein-glycan interactions.

UR - http://www.scopus.com/inward/record.url?scp=80052203282&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052203282&partnerID=8YFLogxK

U2 - 10.1074/jbc.M111.274217

DO - 10.1074/jbc.M111.274217

M3 - Article

VL - 286

SP - 31610

EP - 31622

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 36

ER -