Sushi Peptide Multimer
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Abstract
Endotoxin, also known as lipopolysaccharides (LPS), is the major mediator of septic shock due to Gram-negative bacterial infection. Chemically synthesized S3 peptide, derived from Sushi3 domain of Factor C, which is the endotoxin-sensitive serine protease of the limulus coagulation cascade, binds and neutralizes LPS activity. Fluorescent tagged-S3 is shown to detect LPS-containing bacteria. For large-scale production of S3 and to mimic other pathogen-recognizing molecules, tandem multimers of the S3 gene were constructed and expressed in E. coli. Tetramer of S3 for example is shown to display an enhanced inhibitory effect on LPS-induced activities. An affinity matrix based on tetramer of S3 is also shown to be particularly efficient at removing LPS.
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38 Claims
- 1. A polypeptide comprising more than one S3 peptides.
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14. The polypeptide of olaim 1 any one tagged with a detectable label.
- 15. S3 peptide tagged with a detectable label.
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23. A method of producing a polypeptide having a desired number of S3 peptides, comprising the step of expressing in a host cell DNA encoding a polypeptide which comprises S3 peptides in greater number than the desired number, and wherein at least two of the S3 peptides are separated by a cleavable linking sequence;
- and subjecting the polypeptide to conditions suitable for cleaving the linking sequence to produce the polypeptide having the desired number of S3 peptides while keeping the S3 peptides intact.
- View Dependent Claims (24, 25, 26, 27)
Specification