Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
5
|
| pubmed:dateCreated |
1987-6-18
|
| pubmed:abstractText |
The interaction of the catalytic subunit of bovine cardiac muscle cAMP-dependent protein kinase with N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8), the most potent and selective inhibitor toward cyclic nucleotide-dependent protein kinases in the series of isoquinolinesulfonamide derivatives, was studied. The addition of H-8 protected the catalytic subunit of the enzyme in a dose-dependent manner from irreversible inactivation by the ATP analogue p-fluorosulfonylbenzoyl-5'-adenosine (FSBA). The inactivation followed pseudo-first order kinetics and H-8 reduced the steady state constant of inactivation (Ki) without any effect on the first order rate constant (K3). The quantitative binding of H-8 to the enzyme was measured under conditions of thermodynamic equilibrium using a gel filtration method. The catalytic subunit bound approximately 1 mol of drug/mol of protein with apparent half-maximal binding at 1.0 microM drug, whereas the enzyme irreversibly modified by FSBA did not bind the drug, confirming that the enzyme has no site for H-8 in the catalytic subunit other than the active site. The binding studies also showed that H-8 does not require divalent cations such as Mg2+ to bind to the catalytic subunit of the protein kinase. The binding of H-8 to the active site was characterized using FSBA and other affinity labeling reagents which have been postulated to modify residues at or near the active site of the catalytic subunit. H-8 protected the enzyme against inactivation by FSBA and Cibacron Blue F3GA but did not afford any protection against the covalent modification of 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl), suggesting that the binding site of H-8 does not involve the gamma-subsite of the ATP binding site in the catalytic subunit, since DTNB and NBD-Cl are thought to modify the residues complementary to gamma-phosphate of the ATP molecules.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/4-Chloro-7-nitrobenzofurazan,
http://linkedlifedata.com/resource/pubmed/chemical/5'-(4-fluorosulfonylbenzoyl)adenosin...,
http://linkedlifedata.com/resource/pubmed/chemical/5-carboxymethyl-3-p-tolylthiazolidin...,
http://linkedlifedata.com/resource/pubmed/chemical/Acetophenones,
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine,
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Triphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Affinity Labels,
http://linkedlifedata.com/resource/pubmed/chemical/Caseins,
http://linkedlifedata.com/resource/pubmed/chemical/Cibacron Blue F 3GA,
http://linkedlifedata.com/resource/pubmed/chemical/Dithionitrobenzoic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Histones,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinase Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Thiazolidinediones,
http://linkedlifedata.com/resource/pubmed/chemical/Triazines,
http://linkedlifedata.com/resource/pubmed/chemical/o-Phthalaldehyde
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
May
|
| pubmed:issn |
0026-895X
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
31
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
523-8
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:3574296-4-Chloro-7-nitrobenzofurazan,
pubmed-meshheading:3574296-Acetophenones,
pubmed-meshheading:3574296-Adenosine,
pubmed-meshheading:3574296-Adenosine Triphosphate,
pubmed-meshheading:3574296-Affinity Labels,
pubmed-meshheading:3574296-Animals,
pubmed-meshheading:3574296-Binding Sites,
pubmed-meshheading:3574296-Caseins,
pubmed-meshheading:3574296-Cattle,
pubmed-meshheading:3574296-Dithionitrobenzoic Acid,
pubmed-meshheading:3574296-Histones,
pubmed-meshheading:3574296-Magnesium,
pubmed-meshheading:3574296-Myocardium,
pubmed-meshheading:3574296-Protein Kinase Inhibitors,
pubmed-meshheading:3574296-Structure-Activity Relationship,
pubmed-meshheading:3574296-Thiazolidinediones,
pubmed-meshheading:3574296-Triazines,
pubmed-meshheading:3574296-o-Phthalaldehyde
|
| pubmed:year |
1987
|
| pubmed:articleTitle |
Specific binding of a novel compound, N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8) to the active site of cAMP-dependent protein kinase.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|