Anodonta C reactive protein (CRP), a primeval CRP, was purified from the body fluid of Anodonta Woodiana by affinity chromatography. Purified Anodonta CRP showed a single band in SDS PAGE. Contrasted with the pyroglutamic acid that was found in human or rabbit CRP, the N terminal residue of the Anodonta CRP was Glu. The sequence of the first ten amino acids at the N terminal of the Anodonta CRP was H 2N E T A Y S C I T A V ,which was determined using the modified Edman degradation. The C terminal sequence of the Anodonta CRP had been determined by the carboxypeptidase A degradation. It was L/V S S T Y COOH, different from any other CRP obtained from different species (human or rabbit). The N and C terminal sequences were also found and proved in the tryptic peptides sequences. This Anodonta CRP was digested with trypsin, V8 protease and CNBr respectively. The peptides were separated and collected with the C 18 reversed phase column HPLC and all of the 35 peptides from the HPLC or Sephacryl 200 gel filtration were sequenced. The major structure of the Anodonta CRP was determined by splicing of the sequences from the tryptic peptides, V8 peptides, and CNBr peptides. Both Ca 2+ binding and phosphorylcholine (PC) binding sequences were found in the primary structure. However, except for the two binding sites there was no similar structure to be found between Anodonta CRP and CRP from other species. Additionally, variability was found in the primary sequence of the Anodonta CRP. This phenomenon has been found in the limulus CRP and suggests that the Anodonta CRP is coded by multiple genes.
Key words
Anodonta /
C reactive protein /
Primary structure
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References
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