Detection of the Activities of Multiple Protein Kinases in Lipopolysaccharide-Stimulated Macrophages by Renaturation

Autor:	Masayasu Nakano, Hiroto Shinomiya, Osamu Murakami, Sayaka Utsumi
Rok vydání:	2001
Předmět:	Biochemistry Kinase p38 mitogen-activated protein kinases Mitogen-activated protein kinase biology.protein Phosphorylation Protein phosphorylation General Medicine Biology Receptor Protein kinase A MAPK14
Zdroj:	Journal of Clinical and Experimental Hematopathology. 41:115-118
ISSN:	1880-9952 1346-4280
DOI:	10.3960/jslrt.41.115
Popis:	Past studies of individual kinases have demonstrated that protein phosphorylation plays a crucial role in the intracellular signaling pathway of bacterial lipopolysaccharide (LPS). However, no one has determined how many kinases may be activated collectively by LPS stimulation. We examined the spectrum of protein kinases activated in macrophages in response to LPS. Activity was assessed by a renatuiation method that exploited the eability of proteins denatured with sodium dodcyl sulfate and then blotted onto a membrane to regain enzymatic activity after guanidine treatment. Seven electrophoretically-distinct protein kinases with apparent molecular masses of 78, 74, 62, 59, 58, 52, and 48-kDa were detected in lysates from unstimulated murine peritoneal macrophages. An additional three kinases, with apparent molecular masses of 82, 55, and 46-kDa, were detected when the macrophages were stimulated with LPS. The activation of these protein kinases may be dictated by complex signals that are delivered by receptor complexes, including Toll-like receptor 4. These results should provide a clue to clarifying the pleiotropic action of LPS on macrophages.
Databáze:	OpenAIRE
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