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Titolo:
Characterization of the protein phosphatase 1 catalytic subunit in endothelium: Involvement in contractile responses
Autore:
Verin, AD; Csortos, C; Durbin, SD; Aydanyan, A; Wang, PY; Patterson, CE; Garcia, JGN;
Indirizzi:
Johns Hopkins Univ, Sch Med, Dept Med, Div Pulm & Crit Care Med, Baltimore, MD 21224 USA Johns Hopkins Univ Baltimore MD USA 21224 re Med, Baltimore, MD 21224 USA Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Med, Indianapolis, IN USA Indiana Univ Indianapolis IN USA Med Ctr, Dept Med, Indianapolis, IN USA Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Physiol, Indianapolis, IN USA Indiana Univ Indianapolis IN USA Ctr, Dept Physiol, Indianapolis, IN USA Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Biophys, Indianapolis, IN USA Indiana Univ Indianapolis IN USA Ctr, Dept Biophys, Indianapolis, IN USA
Titolo Testata:
JOURNAL OF CELLULAR BIOCHEMISTRY
fascicolo: 1, volume: 79, anno: 2000,
pagine: 113 - 125
SICI:
0730-2312(2000)79:1<113:COTPP1>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
SMOOTH-MUSCLE MYOSIN; LIGHT-CHAIN PHOSPHORYLATION; CELL GAP FORMATION; ALBUMIN PERMEABILITY; MOLECULAR-CLONING; BARRIER FUNCTION; SKELETAL-MUSCLE; CALYCULIN-A; RAT; EXPRESSION;
Keywords:
microinjection; inhibitor-2; CS1 isoforms in endothelium;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Verin, AD Johns Hopkins Asthma & Allergy Ctr, 5501 Hopkins Bayview Circle,Baltimore, MD 21224 USA Johns Hopkins Asthma & Allergy Ctr 5501 Hopkins Bayview Circle Baltimore MD USA 21224
Citazione:
A.D. Verin et al., "Characterization of the protein phosphatase 1 catalytic subunit in endothelium: Involvement in contractile responses", J CELL BIOC, 79(1), 2000, pp. 113-125

Abstract

We have previously demonstrated the direct involvement of a type 1 Ser/Thrphosphatase (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, 1-2 into bovine pulmonary artery EC and demonstrated both anincrease 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 blotanalysis 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-CS1 alpha and anti-CS1 delta antibodies The anti-CS1 delta antiserum, but not anti-CS1 alpha antiserum cross reacts with the CS1 isoform present in myosin-enriched fraction and CS1 delta 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 CS1 delta-GFP construct in bovine endothelium followed by immunoprecipitation of CS1 with anti-GFP antibody revealed the stable association of CS1 delta with actomyosin complex. Finally, screening of a human EC oligo(dT)-primed cDNA library with a probe encoding a rat CS1 delta cDNA segment yielding several positive clones that encoded the entire CS1 delta open reading frame and partially noncoding regions. Sequence analysis determined a high homology (approximate to 99%) with human CS1 delta derived From a teratocarcinoma cell line. Together, these data suggest that CS1 delta is the major of Prase 1 isoform specifically associated with EC actomyosin complex and which participates in EC barrier regulation. (C) 2000 Wiley-Liss Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 09:06:47