Mapping the heparin‐binding domain of boar spermadhesins

FEBS Letters

Adermann, Knut²  Töpfer-Petersen, Edda¹  Sanz, Libia¹  Dostàlovà, Zuzana¹  Calvete, Juan J.¹  Thole, Hubert H.³ 

[1] Institut für Reproduktionsmedizin, Tierärztliche Hochschule, Bünteweg 15, 30559 Hannover, Germany;Niedersächsisches Institut für Peptid-Forschung GmbH, Hannover, Germany;Max-Planck-Institut für Experimentelle Endokrinologie, Hannover, Germany

关键词: Spermadhesins (boar);    AWN-1;    CUB domain;    Heparin-binding mapping;    Proteolytic protection assay;    Synthetic peptide;   

DOI  :  10.1016/0014-5793(95)01513-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Boar spermadhesins are a group of seminal plasma, heparin-binding proteins which appear to be involved in sperm capacitation and gamete interaction. Using a proteolytic protection assay we have identified regions of AQN-1, AQN-3, PSP-I and AWN which remain attached to a heparin-Sepharose column following in-column digestion of bound spermadhesins with chymotrypsin and elastase. In addition, the complete amino acid sequence of spermadhesin AWN was synthesized as overlapping peptides, and their ability to bind to a heparin-Sepharose column and to inhibit the interaction of soluble heparin with purified ELISA plate-coated AWN was tested. Both approaches gave similar results and as a whole showed that different regions of AWN may converge in its tertiary structure to form a composite heparin-binding site. The conformational heparin-binding surface resides on the GFCC′C″ face of the proposed structural model for AWN and is in an opposite location to the carbohydrate-binding region of the spermadhesin.

【 授权许可】

Unknown