help button home button
AJRCMB
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Verin, A. D.
Right arrow Articles by Garcia, J. G. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Verin, A. D.
Right arrow Articles by Garcia, J. G. N.

Am. J. Respir. Cell Mol. Biol., Volume 19, Number 5, November, 1998 767-776

Biochemical Regulation of the Nonmuscle Myosin Light Chain Kinase Isoform in Bovine Endothelium

Alexander D. Verin, Lydia I. Gilbert-McClain, Carolyn E. Patterson, and Joe G. N. Garcia

Department of Medicine, Physiology and Biophysics, Indiana University School of Medicine, Richard Roudebush Veterans Administration Center, Indianapolis, Indiana

Specific models of vascular permeability are critically dependent on myosin light chain phosphorylation, a reaction catalyzed by a novel high molecular-weight (214 kD) Ca2+/calmodulin (CaM)-dependent myosin light chain kinase (MLCK) isoform recently cloned in human endothelium (Am. J. Respir. Cell Mol. Biol., 1997;16:489-494). To evaluate mechanisms of endothelial cell (EC) barrier dysfunction evoked by the serine protease thrombin, we studied the regulation of the 214-kD EC MLCK isoform expressed in bovine endothelium. The EC MLCK isoform bound biotinylated CaM in a Ca2+-dependent manner and co-immunoprecipitated in a functional complex with myosin, actin, and CaM. Thrombin rapidly increased MLCK activity in concert with time-dependent translocation of the enzyme to the actin cytoskeleton. To evaluate whether EC MLCK activity was regulated by direct phosphorylation, amino acid sequence analysis identified multiple potential EC MLCK sites for Ser/Thr phosphorylation, including highly conserved phosphorylation sites for cyclic adenosine monophosphate-dependent protein kinase A (PKA) adjacent to the CaM-binding region. EC MLCK activity was attenuated by either PKA-mediated MLCK phosphorylation or inhibition of Ser/Thr phosphatase activity (fluoride or calyculin), which significantly increased MLCK phosphorylation while decreasing MLCK activity (3- to 4-fold decrease). In summary, although the EC MLCK isoform exhibits multiple features intrinsic to this family of kinases, thrombin-mediated EC contraction and barrier dysfunction requires increased EC MLCK-actin interaction and MLCK translocation to the cytoskeleton. EC MLCK activity appears to be highly dependent upon the phosphorylation status of this key contractile effector.




This article has been cited by other articles:


Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
C. Csortos, I. Kolosova, and A. D. Verin
Regulation of vascular endothelial cell barrier function and cytoskeleton structure by protein phosphatases of the PPP family
Am J Physiol Lung Cell Mol Physiol, October 1, 2007; 293(4): L843 - L854.
[Abstract] [Full Text] [PDF]

Home page
 GENES CELLSHome page
A. Maeda, Y.-i. Ozaki, S. Sivakumaran, T. Akiyama, H. Urakubo, A. Usami, M. Sato, K. Kaibuchi, and S. Kuroda
Ca-independent phospholipase A2-dependent sustained Rho-kinase activation exhibits all-or-none response.
Genes Cells, September 1, 2006; 11(9): 1071 - 1083.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
L. Gao, A. Grant, I. Halder, R. Brower, J. Sevransky, J. P. Maloney, M. Moss, C. Shanholtz, C. R. Yates, G. U. Meduri, et al.
Novel Polymorphisms in the Myosin Light Chain Kinase Gene Confer Risk for Acute Lung Injury
Am. J. Respir. Cell Mol. Biol., April 1, 2006; 34(4): 487 - 495.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
D. Mehta and A. B. Malik
Signaling Mechanisms Regulating Endothelial Permeability
Physiol Rev, January 1, 2006; 86(1): 279 - 367.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Z. M. Goeckeler and R. B. Wysolmerski
Myosin Phosphatase and Cofilin Mediate cAMP/cAMP-dependent Protein Kinase-induced Decline in Endothelial Cell Isometric Tension and Myosin II Regulatory Light Chain Phosphorylation
J. Biol. Chem., September 23, 2005; 280(38): 33083 - 33095.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
X. Cullere, S. K. Shaw, L. Andersson, J. Hirahashi, F. W. Luscinskas, and T. N. Mayadas
Regulation of vascular endothelial barrier function by Epac, a cAMP-activated exchange factor for Rap GTPase
Blood, March 1, 2005; 105(5): 1950 - 1955.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
A. A. Birukova, F. Liu, J. G. N. Garcia, and A. D. Verin
Protein kinase A attenuates endothelial cell barrier dysfunction induced by microtubule disassembly
Am J Physiol Lung Cell Mol Physiol, July 1, 2004; 287(1): L86 - L93.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
K. G. Birukov, J. R. Jacobson, A. A. Flores, S. Q. Ye, A. A. Birukova, A. D. Verin, and J. G. N. Garcia
Magnitude-dependent regulation of pulmonary endothelial cell barrier function by cyclic stretch
Am J Physiol Lung Cell Mol Physiol, October 1, 2003; 285(4): L785 - L797.
[Abstract] [Full Text] [PDF]

Home page
 Cardiovasc ResHome page
C. Schafer, S. Walther, M. Schafer, L. Dieterich, S. Kasseckert, Y. Abdallah, and H.M. Piper
Inhibition of contractile activation reduces reoxygenation-induced endothelial gap formation
Cardiovasc Res, April 1, 2003; 58(1): 149 - 155.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
I. PETRACHE, K. BIRUKOV, A. L. ZAIMAN, M. T. CROW, H. DENG, R. WADGAONKAR, L. H. ROMER, and J. G. N. GARCIA
Caspase-dependent cleavage of myosin light chain kinase (MLCK) is involved in TNF-{alpha}-mediated bovine pulmonary endothelial cell apoptosis
FASEB J, March 1, 2003; 17(3): 407 - 416.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
J. G. N. GARCIA, P. WANG, K. L. SCHAPHORST, P. M. BECKER, T. BORBIEV, F. LIU, A. BIRUKOVA, K. JACOBS, N. BOGATCHEVA, and A. D. VERIN
Critical involvement of p38 MAP kinase in pertussis toxin-induced cytoskeletal reorganization and lung permeability
FASEB J, July 1, 2002; 16(9): 1064 - 1076.
[Abstract] [Full Text] [PDF]

Home page
 Circ. Res.Home page
S. Y. Yuan, M. H. Wu, E. E. Ustinova, M. Guo, J. H. Tinsley, P. de Lanerolle, and W. Xu
Myosin Light Chain Phosphorylation in Neutrophil-Stimulated Coronary Microvascular Leakage
Circ. Res., June 14, 2002; 90(11): 1214 - 1221.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
P. Wang, A. D. Verin, A. Birukova, L. I. Gilbert-McClain, K. Jacobs, and J. G. N. Garcia
Mechanisms of sodium fluoride-induced endothelial cell barrier dysfunction: role of MLC phosphorylation
Am J Physiol Lung Cell Mol Physiol, December 1, 2001; 281(6): L1472 - L1483.
[Abstract] [Full Text] [PDF]

Home page
 J. Appl. Physiol.Home page
S. M. Dudek and J. G. N. Garcia
Cytoskeletal regulation of pulmonary vascular permeability
J Appl Physiol, October 1, 2001; 91(4): 1487 - 1500.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
B. Nilius and G. Droogmans
Ion Channels and Their Functional Role in Vascular Endothelium
Physiol Rev, October 1, 2001; 81(4): 1415 - 1459.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
A. D. Verin, A. Birukova, P. Wang, F. Liu, P. Becker, K. Birukov, and J. G. N. Garcia
Microtubule disassembly increases endothelial cell barrier dysfunction: role of MLC phosphorylation
Am J Physiol Lung Cell Mol Physiol, September 1, 2001; 281(3): L565 - L574.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
F. Liu, A. D. Verin, T. Borbiev, and J. G. N. Garcia
Role of cAMP-dependent protein kinase A activity in endothelial cell cytoskeleton rearrangement
Am J Physiol Lung Cell Mol Physiol, June 1, 2001; 280(6): L1309 - L1317.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
P. J. Farmer, S. G. Bernier, A. Lepage, G. Guillemette, D. Regoli, and P. Sirois
Permeability of endothelial monolayers to albumin is increased by bradykinin and inhibited by prostaglandins
Am J Physiol Lung Cell Mol Physiol, April 1, 2001; 280(4): L732 - L738.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
A. J. Piekny, A. Wissmann, and P. E. Mains
Embryonic Morphogenesis in Caenorhabditis elegans Integrates the Activity of LET-502 Rho-Binding Kinase, MEL-11 Myosin Phosphatase, DAF-2 Insulin Receptor and FEM-2 PP2c Phosphatase
Genetics, December 1, 2000; 156(4): 1671 - 1689.
[Abstract] [Full Text]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
N. Norwood, T. M. Moore, D. A. Dean, R. Bhattacharjee, M. Li, and T. Stevens
Store-operated calcium entry and increased endothelial cell permeability
Am J Physiol Lung Cell Mol Physiol, November 1, 2000; 279(5): L815 - L824.
[Abstract] [Full Text] [PDF]

Home page
 JCBHome page
Poperechnaya, Varlamova, Lin, J. Stull, and A. Bresnick
Localization and Activity of Myosin Light Chain Kinase Isoforms during the Cell Cycle
J. Cell Biol., October 30, 2000; 151(3): 697 - 708.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
T. Noll, M. Schafer, U. Schavier-Schmitz, and H. M. Piper
ATP induces dephosphorylation of myosin light chain in endothelial cells
Am J Physiol Cell Physiol, September 1, 2000; 279(3): C717 - C723.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
T. Stevens, J. G. N. Garcia, D. M. Shasby, J. Bhattacharya, and A. B. Malik
Mechanisms regulating endothelial cell barrier function
Am J Physiol Lung Cell Mol Physiol, September 1, 2000; 279(3): L419 - L422.
[Abstract] [Full Text] [PDF]

Home page
 Circ. Res.Home page
J. S. Alexander
Rho, Tyrosine Kinase, Ca2+, and Junctions in Endothelial Hyperpermeability
Circ. Res., August 18, 2000; 87(4): 268 - 271.
[Full Text] [PDF]

Home page
 Infect. Immun.Home page
F. Trottein, L. Descamps, S. Nutten, M.-P. Dehouck, V. Angeli, A. Capron, R. Cecchelli, and M. Capron
Schistosoma mansoni Activates Host Microvascular Endothelial Cells To Acquire an Anti-Inflammatory Phenotype
Infect. Immun., July 1, 1999; 67(7): 3403 - 3409.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
J. G. N. Garcia, A. D. Verin, K. Schaphorst, R. Siddiqui, C. E. Patterson, C. Csortos, and V. Natarajan
Regulation of endothelial cell myosin light chain kinase by Rho, cortactin, and p60src
Am J Physiol Lung Cell Mol Physiol, June 1, 1999; 276(6): L989 - L998.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. G. Birukov, C. Csortos, L. Marzilli, S. Dudek, S.-F. Ma, A. R. Bresnick, A. D. Verin, R. J. Cotter, and J. G. N. Garcia
Differential Regulation of Alternatively Spliced Endothelial Cell Myosin Light Chain Kinase Isoforms by p60Src
J. Biol. Chem., March 9, 2001; 276(11): 8567 - 8573.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
E. K. Blue, Z. M. Goeckeler, Y. Jin, L. Hou, S. A. Dixon, B. P. Herring, R. B. Wysolmerski, and P. J. Gallagher
220- and 130-kDa MLCKs have distinct tissue distributions and intracellular localization patterns
Am J Physiol Cell Physiol, March 1, 2002; 282(3): C451 - C460.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Proc. Am. Thorac. Soc. Am. J. Respir. Crit. Care Med.
Copyright © 1998 American Thoracic Society. 		  2009 ATS Conference