Identification of Clostridium perfringens large Sialidase interacting proteins by Yeast Two-Hybrid system and Surface Plasmon Resonance (SPR)
1Institute of Biochemistry, National Yang-Ming University, Taipei 112, Taiwan
2Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan
Sialidases (nanH) are a superfamily of N-acylneuraminate-releasing exoglycosidases found mainly in higher eukaryotes and in some virus, bacteria and protozoans. The previous studies has shown that sialidase could shorten the blood clotting time, in this study, the mechanism of sialidase action involved in shortening blood clotting was investigated. We screened human liver cDNA library with yeast two-hybrid system using large sialidase as bait. Three putative positive clones are related to blood clotting. They are vitronectin, prothrombin and serum albumin. The protein-protein interaction between nanH and each of the three proteins was confirmed with surface plasmon resonance. We proposed a model of large sialidase involved in blood coagulation: the three proteins may stabilize sialidase conformation for easy accessing of sialoglycoconjugates. Desialylation of the sialoglycoconjugates may result in an increase in aggregation rate. We will find out the interaction domain of large sialidase by SPR in the near future.