Characterization of a high-affinity sialic acid-specific CBM40 from Clostridium perfringens and engineering of a divalent form - CEntre de Recherches sur les MAcromolécules Végétales Accéder directement au contenu
Article Dans Une Revue Biochemical Journal Année : 2016

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

Résumé

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.
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hal-03641270 , version 1 (20-04-2022)

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João Ribeiro, William Pau, Carlo Pifferi, Olivier Renaudet, Annabelle Varrot, et al.. Characterization of a high-affinity sialic acid-specific CBM40 from Clostridium perfringens and engineering of a divalent form. Biochemical Journal, 2016, 473 (14), pp.2109-2118. ⟨10.1042/BCJ20160340⟩. ⟨hal-03641270⟩
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