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001-es BibID:	BIBFORM047043
035-os BibID:	PMID:11113114
Első szerző:	Birukov, Konstantin G.
Cím:	Differential regulation of alternatively spliced endothelial cell myosin light chain kinase isoforms by p60(Src) / Konstantin G. Birukov, Csilla Csortos, Lisa Marzilli, Steven Dudek, Shwu-Fan Ma, Anne R. Bresnick, Alexander D. Verin, Robert J. Cotter, Joe G. N. Garcia
Dátum:	2001
ISSN:	0021-9258 1083-351X
Megjegyzések:	The Ca(2+)/calmodulin-dependent endothelial cell myosin light chain kinase (MLCK) triggers actomyosin contraction essential for vascular barrier regulation and leukocyte diapedesis. Two high molecular weight MLCK splice variants, EC MLCK-1 and EC MLCK-2 (210-214 kDa), in human endothelium are identical except for a deleted single exon in MLCK-2 encoding a 69-amino acid stretch (amino acids 436-505) that contains potentially important consensus sites for phosphorylation by p60(Src) kinase (Lazar, V., and Garcia, J. G. (1999) Genomics 57, 256-267). We have now found that both recombinant EC MLCK splice variants exhibit comparable enzymatic activities but a 2-fold reduction of V(max), and a 2-fold increase in K(0.5 CaM) when compared with the SM MLCK isoform, whereas K(m) was similar in the three isoforms. However, only EC MLCK-1 is readily phosphorylated by purified p60(Src) in vitro, resulting in a 2- to 3-fold increase in EC MLCK-1 enzymatic activity (compared with EC MLCK-2 and SM MLCK). This increased activity of phospho-MLCK-1 was observed over a broad range of submaximal [Ca(2+)] levels with comparable EC(50) [Ca(2+)] for both phosphorylated and unphosphorylated EC MLCK-1. The sites of tyrosine phosphorylation catalyzed by p60(Src) are Tyr(464) and Tyr(471) within the 69-residue stretch deleted in the MLCK-2 splice variant. These results demonstrate for the first time that p60(Src)-mediated tyrosine phosphorylation represents an important mechanism for splice variant-specific regulation of nonmuscle MLCK and vascular cell function.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:	Journal of Biological Chemistry. - 276 : 11 (2001), p. 8567-8573. -
További szerzők:	Csortos Csilla (1956-) (biokémikus) Marzilli, Lisa Dudek, Steven Ma, Shwu-Fan Bresnick, Anne R. Verin, Alexander Cotter, Robert J. Garcia, Joe G. N.
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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