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Literature summary for 1.14.13.39 extracted from

  • Tran, Q.K.; Leonard, J.; Black, D.J.; Persechini, A.
    Phosphorylation within an autoinhibitory domain in endothelial nitric oxide synthase reduces the Ca2+ concentrations required for calmodulin to bind and activate the enzyme (2008), Biochemistry, 47, 7557-7566.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Bos taurus

Protein Variants

Protein Variants Comment Organism
additional information deletion of the autoinhibitory domain doubles the maximum calmodulin-dependent enzyme activity and decreases the EC50(Ca2+) values for calmodulin binding and calmodulin-dependent enzyme activation Bos taurus
S617D the phosphomimetic substitution doubles the maximum calmodulin-dependent enzyme activity and decreases the EC50(Ca2+) values for calmodulin binding and enzyme activation compared to the wild type enzyme Bos taurus
S617D/S635D the maximum calmodulin-dependent synthase activity of S617/635D eNOS is about 2fold higher than the wild type activity Bos taurus
S635D the substitution has little or no effect on enzyme activity Bos taurus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required for calmodulin to bind and activate the enzyme, maximum calmodulin-dependent activity is measured at 1.5 mM CaCl2 Bos taurus

Organism

Organism UniProt Comment Textmining
Bos taurus P29473
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA Sepharose column chromatography and 2'-5' ADP Sepharose resin column chromatography Bos taurus

Source Tissue

Source Tissue Comment Organism Textmining

Storage Stability

Storage Stability Organism
-85°C, 50 mM Tris (pH 7.4) with 10% (v/v) glycerol, 100 mM NaCl, 0.2 mM L-arginine, 0.0002 mM 5,6,7,8-tetrahydro-L-biopterin, 0.02 mM dithiothreitol, and 1 mM 2',3'-AMP (mixed isomers), up to 1 month, no detectable loss of catalytic activity Bos taurus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 L-arginine + 3 NADPH + 3 H+ + 4 O2
-
Bos taurus 2 citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
-
?
oxidized cytochrome c + NADPH + O2
-
Bos taurus reduced cytochrome c + NADP+ + H2O
-
?

Synonyms

Synonyms Comment Organism
endothelial nitric oxide synthase
-
Bos taurus
eNOS
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Bos taurus

Cofactor

Cofactor Comment Organism Structure
5,6,7,8-tetrahydro-L-biopterin
-
Bos taurus
Calmodulin maximum calmodulin-dependent activity is measured at 1.5 mM CaCl2, phosphorylation within an autoinhibitory domain in endothelial nitric oxide synthase reduces the Ca2+ concentrations required for calmodulin to bind and activate the enzyme Bos taurus
NADPH
-
Bos taurus