CYCLIC GMP-DEPENDENT PROTEIN-KINASE ACTIVITY IN RAT PULMONARY MICROVASCULAR ENDOTHELIAL-CELLS

被引:22
作者
DIWAN, AH
THOMPSON, WJ
LEE, AK
STRADA, SJ
机构
[1] Depatment of Pharmacology, University of South Alabama, College of Medicine, Mobile
关键词
D O I
10.1006/bbrc.1994.1991
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cyclic GMP-dependent protein kinase (cGPK) activity was determined in rat pulmonary microvascular endothelial cells(RPMVEC) using cGMP-stimulated phosphorylation of BPDEtide and histone F2B substrates in the presence of PKI [peptide inhibitor of cAMP-dependent protein kinase (cAPK)]. RPMVEC cGPK activity was localized to the 100,000 x g cytosolic fraction. The EC(50) for cGMP activation in the presence of PKI was 0.16 mu M and H-89 inhibition under similar conditions showed an IC50 value of 0.16 mu M. Anion-exchange chromatography of RPMVEC and rat lung cytosolic fractions showed separation of the cGMP-dependent from the cGMP-independent protein kinase activity and similar elution conductivities. Further, Western blots of RPMVEC active DEAE-Trisacryl fractions showed immunoreactivity using bovine Type I cGPK antiserum. Preliminary studies reveal six potential substrates phosphorylated by cGPK in RPMVEC. These studies describe an endothelial cell (EC) cGMP-receptor, cGPK, in addition to cGMP-activated (Type II) phosphodiesterase (PDE). (C) 1994 Academic Press, Inc.
引用
收藏
页码:728 / 735
页数:8
相关论文
共 24 条
[1]   AN ACTIVE 20-AMINO-ACID-RESIDUE PEPTIDE DERIVED FROM THE INHIBITOR PROTEIN OF THE CYCLIC AMP-DEPENDENT PROTEIN-KINASE [J].
CHENG, HC ;
VANPATTEN, SM ;
SMITH, AJ ;
WALSH, DA .
BIOCHEMICAL JOURNAL, 1985, 231 (03) :655-661
[2]  
COLBRAN JL, 1992, J BIOL CHEM, V267, P9589
[3]  
DELVECCHIO PJ, 1992, IN VITRO CELL DEV-AN, V28A, P711
[4]   PHARMACOLOGY OF PROTEIN-KINASE INHIBITORS [J].
HIDAKA, H ;
KOBAYASHI, R .
ANNUAL REVIEW OF PHARMACOLOGY AND TOXICOLOGY, 1992, 32 :377-397
[5]  
JACKSON CJ, 1990, J CELL SCI, V96, P257
[6]  
JOYCE NC, 1986, J CYCLIC NUCL PROT, V11, P191
[7]  
KISHI Y, 1992, ADV SEC MESS PHOSPH, V25, P201
[8]   NITRIC OXIDE-INDUCED MICROVASCULAR PERMEABILITY ALTERATIONS - A REGULATORY ROLE FOR CGMP [J].
KUBES, P .
AMERICAN JOURNAL OF PHYSIOLOGY, 1993, 265 (06) :H1909-H1915
[9]  
KUO JF, 1970, J BIOL CHEM, V245, P2493
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+