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

Alexander Dmitriyevich Verin, Virginie Lazar, Ronald J. Torry, Carlos A. Labarrere, Carolyn E. Patterson, Joe G.N. Garcia

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)758-766
Number of pages9
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume19
Issue number5
DOIs
StatePublished - Jan 1 1998
Externally publishedYes

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Myosin-Light-Chain Kinase
Endothelium
Molecular Weight
Molecular weight
Endothelial cells
Protein Isoforms
Endothelial Cells
Muscle
Tissue
Smooth Muscle Myosins
Myosin Light Chains
Phosphorylation
Antibodies
Polymerase chain reaction
RNA-Directed DNA Polymerase
Calmodulin
Reverse Transcriptase Polymerase Chain Reaction
Immunoblotting
Smooth Muscle
Macros

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Expression of a novel high molecular-weight myosin light chain kinase in endothelium. / Verin, Alexander Dmitriyevich; Lazar, Virginie; Torry, Ronald J.; Labarrere, Carlos A.; Patterson, Carolyn E.; Garcia, Joe G.N.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 19, No. 5, 01.01.1998, p. 758-766.

Research output: Contribution to journalArticle

Verin, Alexander Dmitriyevich ; Lazar, Virginie ; Torry, Ronald J. ; Labarrere, Carlos A. ; Patterson, Carolyn E. ; Garcia, Joe G.N. / Expression of a novel high molecular-weight myosin light chain kinase in endothelium. In: American Journal of Respiratory Cell and Molecular Biology. 1998 ; Vol. 19, No. 5. pp. 758-766.
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