Analysis of the effects of Perkinsus marinus proteases on plasma proteins of the Eastern oyster (Crassostrea virginica) and the Pacific oyster (Crassostrea gigas).
We employed two in vitro buffer systems to determine the potential pathogenic effects of Perkinsus marinus serine proteases on the plasma proteins of the eastern oyster (Crassostrea virginica) and the Pacific oyster (Crassostrea gigas). Specifically, this study characterized the oyster plasma protein targets of P. marinus proteases. Additionally, protease-specific inhibitory activity was revealed upon comparison of artificial (PBS) and endogenous (plasma-based) diluents employed during protease digestions. It was found that a C. virginica plasma protein of approximately 35 kDa was eliminated when a standard buffer (PBS) was used as a diluent; however, this protein was preserved when a low-molecular-weight, plasma-based, diluent was used. The results strongly indicate that low-molecular-weight inhibitors of P. marinus proteases are present in oyster plasma. A control (nonparasitic) serine protease, alpha-chymotrypsin, was employed to ascertain the specificity of the protease inhibitors. Although alpha-chymotrypsin possesses ample proteolytic activity for C. virginica plasma proteins, the anti-proteases could specifically inhibit only P. marinus proteases. Such specificity of anti-protease activity is not uncommon among low-molecular-weight serine proteases. The hemolymph target protein was isolated by 2D electrophoresis and isoelectrically isolated for further characterization by N-terminal amino acid sequencing.
Oliver
J L
JL
Department of Environmental Sciences, School of Marine Science, Gloucester Point, Virginia 23062, USA.
Lewis
T D
TD
Faisal
M
M
Kaattari
S L
SL
D000818:Animals; D016782:Apicomplexa / Q000201:enzymology*; D001798:Blood Proteins / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D008970:Molecular Weight; D010125:Ostreidae / Q000378:metabolism / Q000469:parasitology*; D011480:Protease Inhibitors / Q000097:blood
D001798:Blood Proteins; D011480:Protease Inhibitors; D010450:Endopeptidases