Autologous extracellular cytochrome c is an endogenous ligand for leucine-rich alpha2-glycoprotein and beta-type phospholipase A2 inhibitor.

Beta-type phospholipase A(2) inhibitory protein (PLIbeta) from the serum of the venomous snake Gloydius brevicaudus neutralizes basic phospholipase A(2) (PLA(2)) from its own venom, and it has 33% sequence homology with human leucine-rich alpha(2)-glycoprotein (LRG), which has been recently reported to bind cytochrome c (Cyt c) (Cummings, C., Walder, J., Treeful, A., and Jemmerson, R. (2006) Apoptosis 11, 1121-1129). In the present study, PLIbeta was found to bind Cyt c. The interactions of LRG and PLIbeta with Cyt c were compared by surface plasmon resonance analysis. Human LRG bound horse and snake Cyt c with dissociation constants of 1.58 x 10(-13) M and 1.65 x 10(-10) M, respectively, but did not bind yeast Cyt c, while G. brevicaudus PLIbeta bound horse, snake, and yeast Cyt c with dissociation constants of 1.05 x 10(-10) M, 2.37 x 10(-12) M, and 1.67 x 10(-6) M, respectively. On the other hand, LRG did not show any PLA(2) inhibitory activity and did not bind G. brevicaudus basic PLA(2), whereas PLIbeta bound the basic PLA(2) with a dissociation constant of 1.21 x 10(-9) M, which is smaller than those with the Cyt c described above. The PLA(2) inhibitory activity of PLIbeta was also found to be suppressed by the binding of Cyt c to PLIbeta. These results suggest that autologous Cyt c is an endogeneous ligand for LRG and PLIbeta and that these serum proteins neutralize the autologous Cyt c released from the dead cells.

Up-regulation of the expression of leucine-rich alpha(2)-glycoprotein in hepatocytes by the mediators of acute-phase response.

Leucine-rich alpha(2)-glycoprotein (LRG) is a plasma protein in which leucine-rich repeats (LRRs) were first discovered. Although the physiological function of LRG is not known, increases in the serum level of LRG have been reported in various diseases. In this study, we found that LRG was induced by recombinant human IL-6 in human hepatoma HepG2 cells. The induction of LRG by IL-6 was up-regulated synergistically with either IL-1beta or TNFalpha in a pattern similar to those for type 1 acute-phase proteins. We also found that lipopolysaccharide (LPS) administered intraperitoneally to mice enhanced dose-dependently the expression of LRG mRNA in the liver as well as those for mouse major acute-phase proteins. These results strongly suggest that LRG was a secretory type 1 acute-phase protein whose expression was up-regulated by the mediator of acute-phase response.