Conduct unbecoming: C-reactive protein interactions with a broad range of protein molecules.

BACKGROUND: C-reactive protein (CRP), a pentamer composed of five identical 23-kd subunits, is a member of a highly conserved family of proteins known as pentraxins. CRP has been recognized as a risk factor for the development of both the native and transplant-associated forms of atherosclerosis. Understanding the biology of CRP may be relevant to understanding atherosclerosis development and progression. METHODS: Using Western-blotting techniques, we examined the interactions between native, monomeric and mutationally and chemically modified CRP and a variety of antibodies, monoclonal and polyclonal. RESULTS: CRP in its denatured monomeric form, but not in its native pentameric conformation, associates promiscuously with IgG molecules, including normal human IgG, as well as with a number of other proteins. This behavior is intrinsic to CRP and is not noted with other pentraxins such as serum amyloid P component or the long pentraxin, PTX3. Monomeric CRP co-localizes with vitronectin in human heart tissue sections. CONCLUSIONS: We present these findings as cautionary advice, to indicate that characterization of monomeric CRP can be complicated by the propensity of the molecule to interact with a variety of immunoglobulins and other proteins. We also suggest that it is possible that such interactions could serve to eliminate excess of monomeric CRP and/or to scavenge altered, damaged and denatured proteins. These reactivities may be part of a regulatory mechanism to limit inflammation in the arterial wall.

SU1498, an inhibitor of vascular endothelial growth factor receptor 2, causes accumulation of phosphorylated ERK kinases and inhibits their activity in vivo and in vitro.

SU1498, an inhibitor of vascular endothelial growth factor receptor 2, has been used successfully to study the physiological manifestations of receptor functions. Here we report that in addition to its anti-receptor activity, SU1498 stimulates accumulation of phosphorylated ERKs in human umbilical vein endothelial cells and in human aortic endothelial cells in a manner that is dependent on the functioning of the upstream components of the MAPK pathway, B-Raf, and MEK kinases. The enhanced accumulation of phospho-ERKs is observed only in cells that have been stimulated with sphingosine 1-phosphate or protein growth factors; SU1498 by itself is ineffective. We show that the inhibitor acts by blocking the kinase activity of phospho-ERK both in a direct assay and in immunoprecipitates from cells treated with the compound. The data reveal a novel and unique way in which MAPK signaling pathway may be blocked in human endothelial cells.