Expression of a novel high molecular-weight myosin light chain kinase in endothelium.

TitleExpression of a novel high molecular-weight myosin light chain kinase in endothelium.
Publication TypeJournal Article
Year of Publication1998
AuthorsVerin AD, Lazar V, Torry RJ, Labarrere CA, Patterson CE, Garcia JG
JournalAm J Respir Cell Mol Biol
Volume19
Issue5
Pagination758-66
Date Published1998 Nov
ISSN Number1044-1549
KeywordsAnimals, Cattle, Cells, Cultured, Endothelium, Vascular, Gene Expression Regulation, Humans, Immunoblotting, Immunohistochemistry, Isoenzymes, Muscle, Smooth, Vascular, Myocardium, Myosin-Light-Chain Kinase, Phosphorylation, Rats
Abstract

<p>Myosin light chain phosphorylation results in cellular contraction and is a critical component of agonist-mediated endothelial cell (EC) junctional gap formation and permeability. We have shown that this reaction is catalyzed by a novel high molecular-weight Ca2+/calmodulin-dependent nonmuscle myosin light chain kinase (MLCK) isoform recently cloned in human endothelium (Am. J. Respir. Cell Mol. Biol., 1997;16:489-494). To characterize EC MLCK expression further in cultured and adult tissues, we employed immunoblotting techniques and reverse transcriptase-polymerase chain reaction to demonstrate that freshly isolated and cultured human macro- and microvascular EC express only the EC MLCK isoform (214 kD), which is distinct from smooth-muscle MLCK isoforms (130 to 150 kD). Immunocytochemical studies demonstrated the presence of the high molecular-weight MLCK isoform in adult human cardiac endothelium using anti-MLCK antibodies, which preferentially recognize the high molecular-weight EC MLCK isoform. Monitoring of MLCK expression in different cell types with antibodies generated against a unique human EC MLCK N-terminal sequence revealed a high level of expression of the 214-kD enzyme in endothelium, minimal level of expression in smooth muscle, and no expression in skeletal muscle. These data suggest that the novel 214-kD kinase, the only MLCK isoform found in endothelium, may be preferentially expressed in this nonmuscle tissue.</p>

DOI10.1165/ajrcmb.19.5.3125
Alternate JournalAm. J. Respir. Cell Mol. Biol.
PubMed ID9806740
Grant ListHL50533 / HL / NHLBI NIH HHS / United States
HL57462 / HL / NHLBI NIH HHS / United States
HL58064 / HL / NHLBI NIH HHS / United States