Characterization of a high-affinity sialic acid-specific CBM40 from Clostridium perfringens and engineering of a divalent form

João P Ribeiro, William Pau, Carlo Pifferi, Olivier Renaudet, Annabelle Varrot, Lara K Mahal, Anne Imberty

Research output: Contribution to journalArticlepeer-review

Abstract

CBMs (carbohydrate-binding modules) are a class of polypeptides usually associated with carbohydrate-active enzymatic sites. We have characterized a new member of the CBM40 family, coded from a section of the gene NanI from Clostridium perfringens Glycan arrays revealed its preference towards α(2,3)-linked sialosides, which was confirmed and quantified by calorimetric studies. The CBM40 binds to α(2,3)-sialyl-lactose with a Kd of ∼30 μM, the highest affinity value for this class of proteins. Inspired by lectins' structure and their arrangement as multimeric proteins, we have engineered a dimeric form of the CBM, and using SPR (surface plasmon resonance) we have observed 6-11-fold binding increases due to the avidity affect. The structures of the CBM, resolved by X-ray crystallography, in complex with α(2,3)- or α(2,6)-sialyl-lactose explain its binding specificity and unusually strong binding.

Original languageEnglish (US)
Pages (from-to)2109-18
Number of pages10
JournalBiochemical Journal
Volume473
Issue number14
DOIs
StatePublished - Jul 15 2016

Keywords

  • Journal Article

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