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Ligation of cell surface-associated glucose-regulated protein 78 by receptor-recognized forms of alpha 2-macroglobulin: activation of p21-activated protein kinase-2-dependent signaling in murine peritoneal macrophages.

Misra UK, Sharma T, Pizzo SV. Ligation of cell surface-associated glucose-regulated protein 78 by receptor-recognized forms of alpha 2-macroglobulin: activation of p21-activated protein kinase-2-dependent signaling in murine peritoneal macrophages. Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA.  

Previous studies of the plasma proteinase inhibitor alpha2-macroglobulin(alpha2M) demonstrated that alpha2M-proteinase complexes (alpha2M*) modulateimmune responses and promotes macrophage locomotion and chemotaxis. Alpha2M*binds to cell surface-associated glucose-regulated protein 78 (GRP78), whichactivates downstream signaling events. The role of p21-activated protein kinase-1and -2 (PAK-1 and -2) in promoting cellular motility is well documented. In thecurrent study, we examined the ability of alpha2M* to activate PAK-1 and PAK-2.Upon macrophage stimulation with alpha2M*, PAK-2 is autophosphorylated, resultingin increased kinase activity; however, PAK-1 is negligibly affected.
Alpha2M*-stimulated macrophages showed a marked elevation in the levels of Rac x GTP. Receptor tyrosine phosphorylation upon binding of alpha2M* to GRP78,recruits PAK-2 to the plasma membrane via the adaptor protein NCK. Consistentwith this hypothesis, silencing of GRP78 gene expression greatly attenuated thelevels of membrane-associated PAK-2 and NCK. PAK-2 activity was markedlydecreased by inhibition of tyrosine kinases and PI3K before alpha2M* stimulation.We further demonstrate that phosphorylation of Lin-11, Isl-1, Mec-3 (LIM) kinase and cofilin is promoted by treating macrophages with alpha2M*. Thus, alpha2M*regulates activation of the PAK-2-dependent motility mechanism in these cells.
PMID:
16081825 [PubMed - indexed for MEDLINE]