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001-es BibID:	BIBFORM029024
Első szerző:	Kim, Kyung-mi
Cím:	Molecular characterization of myosin phosphatase in endothelium / Kyung-mi Kim, Csilla Csortos, Istvan Czikora, David Fulton, Nagavedi S. Umapathy, Gabor Olah, Alexander D. Verin
Dátum:	2012
ISSN:	0021-9541
Megjegyzések:	The phosphorylation status of myosin light chain (MLC) is regulated by both MLC kinases and type 1 Ser/Thr phosphatase (PPase 1), MLC phosphatase (MLCP) activities. The activity of the catalytic subunit of MLCP (CS1β) towards myosin depends on its associated regulatory subunit, namely myosin PPase targeting subunit 1 (MYPT1). Our previously published data strongly suggested the involvement of MLCP in endothelial cell (EC) barrier regulation. In this study, our new data demonstrate that inhibition of MLCP by either CS1β or MYPT1 siRNA-based depletion results in significant attenuation of purine nucleotide (ATP and adenosine)-induced EC barrier enhancement. Consistent with the data, thrombin-induced EC F-actin stress fiber formation and permeability increase were attenuated by the ectopic expression of constitutively active (C/A) MYPT1. The data demonstrated for the first time direct involvement of MLCP in EC barrier enhancement/protection. Cloning of MYPT1 in human pulmonary artery EC (HPAEC) revealed the presence of two MYPT1 isoforms, long and variant 2 (V2) lacking 56 amino acids from 553 to 609 of human MYPT1 long, which were previously identified in HeLa and HEK 293 cells. Our data demonstrated that in Cos-7 cells ectopically expressed EC MYPT1 isoforms co-immunoprecipitated with intact CS1β suggesting the importance of PPase 1 activity for the formation of functional complex of MYPT1/CS1β. Interestingly, MYPT1 V2 shows decreased binding affinity compared to MYPT1 long for radixin (novel MLCP substrate and a member of ERM family proteins). These results suggest functional difference between EC MYPT1 isoforms in the regulation of MLCP activity and cytoskeleton. J. Cell. Physiol. 227: 1701-1708, 2012. © 2011 Wiley Periodicals, Inc.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban egyetemen (Magyarországon) készült közlemény
Megjelenés:	Journal of Cellular Physiology. - 227 : 4 (2012), p. 1701-1708. -
További szerzők:	Fulton, David Umapathy, Nagavedi S. Oláh Gábor Verin, Alexander Csortos Csilla (1956-) (biokémikus) Czikora István (1979-) (vegyész, biokémikus)
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