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Recombinant Factor C from Carcinoscorpius Rotundicauda Binds Endotoxin

  • A. W. M. Pui
  • S. D. Roopashree
  • B. Ho
  • J. L. Ding
Chapter

Abstract

The coagulation cascade in the circulating amoebocytes of the Singapore horseshoe crab, Carcinoscorpius rotundicauda, plays a vital role in the crab’s defence against invading Gram negative bacteria present in its habitat. Factor C, a serine protease zymogen, is activated by Gram negative bacterial endotoxin or lipopolysaccharide (LPS) to initiate this coagulation cascade. The Limulus gelation assay for detection of endotoxin is based on the affinity of the endotoxin-binding domain of Factor C for LPS [Ho, 1983; Levin, 1985]. Consequently, C. rotundicauda Factor C cDNA has been cloned and expressed in the methylotrophic yeast, Pichia pastoris, to produce recombinant Factor C (rFC) [Roopashree et al., 1996] which will serve as an alternative source of the enzyme to the conventional amoebocyte lysate. This paper describes the biological activity of full-length rFC viz its immunoreactivity by Western detection and particularly, the functionality of its endotoxinbinding domain to bind specifically to LPS and lipid A.

Keywords

Pichia Pastoris Coagulation Cascade Methylotrophic Yeast Yeast Lysate Yeast Cell Lysate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • A. W. M. Pui
    • 1
  • S. D. Roopashree
    • 1
  • B. Ho
    • 2
  • J. L. Ding
    • 1
  1. 1.School of Biological SciencesNational University of SingaporeKent RidgeSingapore
  2. 2.Department of MicrobiologyNational University of SingaporeKent, RidgeSingapore

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