Affinity and kinetic analysis of the bovine plasma C‐type lectin collectin‐43 (CL‐43) interacting with mannan

FEBS Letters

Fischer, Per B.²  Holmskov, Uffe³  Højrup, Peter¹  Rothmann, Anette¹ 

[1] Institute of Molecular Biology, University of Odense, DK-5000 Odense, Denmark;MRC Immunochemistry Unit, The Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK;Department of Medical Microbiology, Institute of Medical Biology, University of Odense, DK-5000 Odense, Denmark

关键词: Collectin-43;    C-type lectin;    Affinity;    Kinetics;    BIAcore;   

DOI  :  10.1016/0014-5793(96)00915-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Collectins are C-type lectins which have been implied to play an important role in the innate immune defence against microorganisms. The critical discriminatory event in the opsonization of microorganisms by collectins is the interaction of the C-type lectin domain with microbial carbohydrates. Surface plasmon resonance measurements allow for quantitative real-time measurements of binding interaction between immobilized carbohydrate and unlabelled lectin in solution. Binding analysis were carried out with purified collectin-43 (CL-43) which structurally is the simplest collectin consisting of only three polypeptides each terminating in a C-type lectin domain. The target was immobilized yeast mannan. The molecular mass of native CL-43 was determinated by mass spectroscopy to 99.8 kDa. The dissociation rate (k _diss) of the C-type lectin-carbohydrate binding was fast (1.19–1.36 × 10⁻² second⁻¹), and the association rate (k _ass) was 4.37–5.07 × 10⁵ M^{−1] second−1} . The equilibrium constant for dissociation (K _d) was 2.68–2.72 × 10⁻⁸ M.

【 授权许可】

Unknown