Intracellular serine-protease zymogen, factor C, from horseshoe crab hemocytes. Its activation by synthetic lipid A analogues and acidic phospholipids.
An intracellular clotting factor, factor C, found in the horseshoe crab hemocytes is a lipopolysaccharide-sensitive serine-protease zymogen, which participates in the initiation of the hemolymph clotting system [T. Nakamura et al. (1986) Eur. J. Biochem. 154, 511-521]. The subsequent study of this zymogen, using various synthetic lipid A analogues, revealed that the zymogen factor C is rapidly activated by acylated (beta 1-6)-D-glucosamine disaccharide bisphosphate (synthetic Escherichia coli-type lipid A), and the corresponding 4'-monophosphate analogues. However, the corresponding non-phosphorylated lipid A did not activate factor C, indicating that a phosphate ester group linked with the (beta 1-6)-D-glucosamine disaccharide backbone is required for the zymogen activation. During these studies we also found that the zymogen factor C is significantly activated by acidic phospholipids, such as phosphatidylinositol, phosphatidylglycerol and cardiolipin, but not at all by neutral phospholipids. The rate of this activation, however, was affected markedly by ionic strength in the reaction mixture, although such an effect was not observed in the lipid-A-mediated activation of factor C. A variety of negatively charged surfaces, such as sulfatide, dextran sulfate and ellagic acid, which are known as typical initiators for activation of the mammalian intrinsic clotting system, did not show any effect on the zymogen factor C activation. These results suggest that lipid A is the most effective trigger to initiate the activation of the horseshoe crab hemolymph clotting system.
Nakamura
T
T
Department of Biology, Faculty of Science, Kyushu University, Japan.
Tokunaga
F
F
Morita
T
T
Iwanaga
S
S
Kusumoto
S
S
Shiba
T
T
Kobayashi
T
T
Inoue
K
K
D000818:Animals; D060829:Arthropod Proteins; D001773:Blood Cells / Q000201:enzymology*; D001779:Blood Coagulation Factors / Q000302:isolation & purification*; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000302:isolation & purification*; D006434:Hemocytes / Q000201:enzymology*; D006737:Horseshoe Crabs / Q000201:enzymology*; D008050:Lipid A / Q000031:analogs & derivatives* / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D012697:Serine Endopeptidases / Q000302:isolation & purification*
D060829:Arthropod Proteins; D001779:Blood Coagulation Factors; D004792:Enzyme Precursors; D008050:Lipid A; D008070:Lipopolysaccharides; D010743:Phospholipids; D012697:Serine Endopeptidases; C057151:Limulus clotting factor C