Intracellular interaction of myosin light chain kinase with macrophage migration inhibition factor (MIF) in endothelium
- 18 April 2005
- journal article
- research article
- Published by Wiley in Journal of Cellular Biochemistry
- Vol. 95 (4), 849-858
- https://doi.org/10.1002/jcb.20472
Abstract
The endothelial cell Ca2+/calmodulin (CaM)-dependent myosin light chain kinase isoform (EC MLCK) is a multifunctional contractile effector involved in vascular barrier regulation, leukocyte diapedesis, apoptosis, and angiogenesis. The EC MLCK isoform and its splice variants contain a unique N-terminal sequence not present in the smooth muscle MLCK isoform (SM MLCK), which allows novel upregulation of MLCK activation by signaling cascades including p60src. The yeast two-hybrid assay system using the entire EC MLCK1 N-terminus (922 aa) as bait, identified additional stable MLCK binding partners including the 12 KDa macrophage migration inhibitory factor (MIF). This finding was confirmed by cross immunoprecipitation assays under non-denaturing conditions and by GST pull down experiments using GST-N-terminal MLCK (#1–923) and MLCK N-terminal deletion mutants in TNFα- and thrombin-stimulated endothelium. This EC MLCK–MIF interaction was shown biochemically and by immunofluorescent microscopy to be enhanced in TNFα- and thrombin-stimulated endothelium, both of which induce increased MLCK activity. Thrombin induced the colocalization of an epitope-tagged, full-length MIF fusion protein with phosphorylated MLC along peripheral actin stress fibers. Together these studies suggest that the novel interaction between MIF and MLCK may have important implications for the regulation of both non-muscle cytoskeletal dynamics as well as pathobiologic vascular events that involve MLCK.Keywords
This publication has 43 references indexed in Scilit:
- Macrophage Migration Inhibitory Factor Is Induced by Thrombin and Factor Xa in Endothelial CellsPublished by Elsevier BV ,2004
- Caspase‐dependent cleavage of myosin light chain kinase (MLCK) is involved in TNF‐α‐mediated bovine pulmonary endothelial cell apoptosisThe FASEB Journal, 2003
- Mutation analysis of the non‐muscle myosin light chain kinase (MLCK) deletion constructs on CV1 fibroblast contractile activity and proliferationJournal of Cellular Biochemistry, 2002
- Myosin II Light Chain Phosphorylation Regulates Membrane Localization and Apoptotic Signaling of Tumor Necrosis Factor Receptor-1Published by Elsevier BV ,2001
- Differential Regulation of Alternatively Spliced Endothelial Cell Myosin Light Chain Kinase Isoforms by p60SrcPublished by Elsevier BV ,2001
- A Proinflammatory Cytokine Inhibits P53 Tumor Suppressor ActivityThe Journal of Experimental Medicine, 1999
- IDENTIFICATION OF MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IN HUMAN VASCULAR ENOTHELIAL CELLS AND ITS INDUCTION BY LIPOPOLYSACCHARIDECytokine, 1998
- Thrombin-induced phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein in bovine pulmonary artery endothelial cellsJournal of Cellular Physiology, 1996
- The role of microtubules in the response of macrophages to MIFCellular Immunology, 1979
- The Mechanism of Action of Macrophage Migration Inhibitory Factor (MIF)International Archives of Allergy and Immunology, 1973