REGULATION OF ENDOTHELIAL CELL CONTRACTION

  • Langan, George Parsons (PI)
  • Ratneshwar, Lal (PI)
  • Garcia, Joe G N (PI)
  • Birukov, Konstantin (PI)
  • Natarajan, Viswanathan (PI)
  • Salgia, Ravi (PI)
  • English, Denis Kelvin (PI)
  • Patterson, Carolyn (PI)
  • Romer, Lewis (PI)
  • Crow, Michael T. (PI)
  • Hassoun, Paul (PI)
  • Verin, Alexander Dmitriyevich (PI)
  • Ratneshwar, Lal (PI)
  • Garcia, Joe G N (PI)
  • Langan, George Parsons (PI)
  • Natarajan, Viswanathan (PI)
  • Salgia, Ravi (PI)
  • Patterson, Carolyn (PI)
  • English, Denis Kelvin (PI)
  • Romer, Lewis (PI)
  • Crow, Michael T. (PI)
  • Hassoun, Paul (PI)
  • GARCIA, JOE (PI)
  • GARCIA, JOE (PI)
  • Natarajan, Viswanathan (PI)
  • Garcia, Joe G N (PI)
  • Patterson, Carolyn (PI)
  • English, Denis Kelvin (PI)
  • Natarajan, Viswanathan (PI)
  • GARCIA, JOE (PI)
  • Natarajan, Viswanathan (PI)
  • CROW, MICHAEL (PI)
  • Birukov, Konstantin (PI)
  • Salgia, Ravi (PI)
  • Langan, George Parsons (PI)
  • LAL, RATNESHWAR (PI)
  • GARCIA, JOE (PI)
  • Birukov, Konstantin (PI)
  • LAL, RATNESHWAR (PI)

Project: Research project

Project Details

Description

Loss of the semi-selective endothelial cell barrier increases vascular
permeability, produces life-threatening pulmonary edema, and is a cardinal
feature of inflammatory lung injury. The regulatory mechanisms underlying
endothelial cell barrier dysfunction, however, are poorly understood. Our
studies support our original working model that barrier dysfunction evoked
by the multifunctional serine protease, thrombin, results from endothelial
cell contraction, gap formation, and increased paracellular fluid and
proteins transport. Our data indicate that thrombin-induced endothelial
contractile events are critically dependent upon activation of a novel,
high molecular weight, Ca2+/calmodulin-dependent myosin light chain kinase
(MLCK). We have recently cloned this unique non-muscle MLCK isoform and
hypothesize that this enzyme provides the molecular machinery for force
generation and endothelial cell barrier dysfunction. In SA#1 we will
utilize standard molecular biological techniques to express and purify
endothelial cell MLCK in order to more closely examine its enzymatic
characteristics. Our findings indicate thrombin-mediated MLCK activation,
MLCK phosphorylation and barrier function are under key regulation by
cAMP-dependent protein kinase A and protein kinase C, with possibly
contributory regulation by Ca2+ and calmodulin availability) are poorly
understood, SA#2 will explore the role of MLCK
phosphorylation/dephosphorylation in the regulation of MLCK activities.
Finally, both the endothelial cell and smooth muscle MLCK gene contain
specific domains whose functions is likely involved in contractile protein
binding and possibly myosin filament stabilization. Based upon our
preliminary characterization of a functional complex of MLCK and MLCK-
binding proteins, SA#3 will identify relevant contractile and signaling
effectors which bind MLCK, and determine both the sites and functional
consequence of MLCK binding. These studies will define the biochemical
basis of endothelial cell actomyosin activation and elucidate the role of
contractile proteins in control of barrier function. This information may
mead to novel therapies designed to preserve the endothelial cell barrier,
reduced alveolar flooding, and limit the patient morbidity characteristic
of acute lung injury syndromes.
StatusFinished
Effective start/end date2/1/981/31/14

Funding

  • National Heart, Lung, and Blood Institute

ASJC

  • Medicine(all)

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