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001-es BibID:	BIBFORM047044
035-os BibID:	PMID:10906760
Első szerző:	Verin, Alexander
Cím:	Characterization of the protein phosphatase 1 catalytic subunit in endothelium : involvement in contractile responses / Alexander D. Verin, Csilla Csortos, Steve D. Durbin, Antonina Aydanyan, Peiyi Wang, Carolyn E. Patterson, Joe G. N. Garcia
Dátum:	2000
ISSN:	0730-2312
Megjegyzések:	We have previously demonstrated the direct involvement of a type 1 Ser/Thr phosphatase (PPase 1) in endothelial cell (EC) barrier regulation [Am. J. Physiol. 269:L99-L108, 1995]. To further extend this observation, we microinjected either the Ser/Thr PPase inhibitor, calyculin, or the PPase 1 inhibitory protein, I-2 into bovine pulmonary artery EC and demonstrated both an increase in F-actin stress fibers and a shift from a regular polygonal shape to a spindle shape with gaps apparent at the cell borders. Northern blot analysis with specific cDNA probes revealed the presence of three major PPase 1 catalytic subunit (CS1) isoforms (alpha, delta, and gamma) in human and bovine EC. To characterize the myosin-associated EC CS1 isoform, myosin-enriched bovine EC fraction was screened with anti-CS1alpha and anti-CS1delta antibodies The anti-CS1delta antiserum, but not anti-CS1alpha antiserum cross reacts with the CS1 isoform present in myosin-enriched fraction and CS1delta was found in stable association with EC myosin/myosin light chain kinase (MLCK) complex in MLCK immunoprecipitates under nondenaturing conditions. Consistent with these data, overexpression of CS1delta-GFP construct in bovine endothelium followed by immunoprecipitation of CS1 with anti-GFP antibody revealed the stable association of CS1delta with actomyosin complex. Finally, screening of a human EC oligo(dT)-primed cDNA library with a probe encoding a rat CS1delta cDNA segment yielding several positive clones that encoded the entire CS1delta open reading frame and partially noncoding regions. Sequence analysis determined a high homology ( approximately 99%) with human CS1delta derived from a teratocarcinoma cell line. Together, these data suggest that CS1delta is the major of PPase 1 isoform specifically associated with EC actomyosin complex and which participates in EC barrier regulation.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:	Journal of Cellular Biochemistry. - 79 : 1 (2000), p. 113-125. -
További szerzők:	Csortos Csilla (1956-) (biokémikus) Durbin, Steve D. Aydanyan, Antonina Wang, Peiyi Patterson, Carolyn E. Garcia, Joe G. N.
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat
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