Purification of a protease inhibitor which controls prophenoloxidase activation in hemolymph of Locusta migratoria (insecta).
A protein which inhibits the prophenoloxidase----phenoloxidase (EC 1.14.18.1) proteolytic activation in hemocyte extracts of Locusta migratoria was isolated from the plasma of the same insect and partially characterized. It shows a molecular weight of 14,000, an inhibiting activity toward the cascade system in the insect hemocytes, which resulted in a lower production of phenoloxidase, a key enzyme for the defence mechanism in arthropods. To identify the specificity of the Locusta inhibitor and consequently the specificity of its target enzyme, inhibitory tests were performed against a number of known serine-proteases. A strong in vitro inhibiting activity toward chymotrypsin and, to a lesser extent, toward human leukocyte elastase was present, while trypsin, Carlsberg subtilisin, human thrombin and pancreatic elastase failed to react. The lack of trypsin inhibition by the isolated inhibitor suggested that the trypsin-catalysed activation of the system in the hemocyte extract takes place under different controls or at an earlier stage of the cascade. The N-terminal sequence of the inhibitor reveals that this molecule is different from the protease inhibitors isolated from other arthropods.
Brehélin
M
M
Laboratoire de Pathologie Comparée, INRA-CNRS (URA 1184), Montpellier, France.
Boigegrain
R A
RA
Drif
L
L
Coletti-Previero
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D004156:Catechol Oxidase / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000187:drug effects*; D006110:Grasshoppers; D006458:Hemolymph / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D013379:Substrate Specificity
D004792:Enzyme Precursors; D011480:Protease Inhibitors; C038451:pro-phenoloxidase; D004156:Catechol Oxidase; D002918:Chymotrypsin