Literature summary for 1.14.15.4 extracted from

Strushkevich, N.; Gilep, A.A.; Shen, L.; Arrowsmith, C.H.; Edwards, A.M.; Usanov, S.A.; Park, H.W.
Structural insights into aldosterone synthase substrate specificity and targeted inhibition (2013), Mol. Endocrinol., 27, 315-324.

Search for more literature for 1.14.15.4

Application

Application	Comment	Organism
drug development	the enzyme is a target for development of selective and potent CYP11B2 inhibitors	Homo sapiens
medicine	blocking of aldosterone synthesis by mediating aldosterone synthase activity is a possible pharmacological therapy for hypertension	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
expression of mature form of CYP11B1 protein without the mitochondrial targeting peptide and with N-terminal tag MAKKTSS added to the 26th residue of CYP11B1, and with C-terminal His6-tag, coexpression with GroEL/ES in Escherichia coli strain JM109	Homo sapiens
expression of mature form of CYP11B2 protein without the mitochondrial targeting peptide and with N-terminal tag MAKKTSS added to the 30th residue of CYP11B2, and with C-terminal His6-tag, coexpression with GroEL/ES in Escherichia coli strain JM109	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant N-terminally MAKKTSS-tagged and C-terminally His6-tagged CYP11B2 without mitochondrial targeting peptide complexed with substrate or inhibitor, hanging drop vapor diffusion method, 14 mg/ml protein in 50 mM potassium phosphate, pH 7.4, containing 500 mM NaCl, 0.5% CHAPS, 20% glycerol, and 0.05 mM 11-deoxycortisosterone or 0.05 mM fadrozole, mixing with reservoir solution containing 10% PEG 4000, 0.2 M lithium sulfate, and 0.1 M Tris, pH 8.5, X-ray diffraction structure determination and analysis at 2.5-2.7 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant N-terminally MAKKTSS-tagged and C-terminally His6-tagged CYP11B2 without mitochondrial targeting peptide complexed with substrate or inhibitor, hanging drop vapor diffusion method, 14 mg/ml protein in 50 mM potassium phosphate, pH 7.4, containing 500 mM NaCl, 0.5% CHAPS, 20% glycerol, and 0.05 mM 11-deoxycortisosterone or 0.05 mM fadrozole, mixing with reservoir solution containing 10% PEG 4000, 0.2 M lithium sulfate, and 0.1 M Tris, pH 8.5, X-ray diffraction structure determination and analysis at 2.5-2.7 A resolution

Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
fadrozole	isoform-selective inhibitor binding. Fadrozole binds to aldosterone synthase in the R-configuration, using part of the active site cavity pointing toward the egress channel	Homo sapiens
additional information	structure homology modelling for inhibitor design, overview	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
11-deoxycorticosterone + reduced adrenodoxin + O2	Homo sapiens	-	corticosterone + oxidized adrenodoxin + H2O	-	?
corticosterone + reduced adrenodoxin + O2	Homo sapiens	-	18-hydroxycorticosterone + oxidized adrenodoxin + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P15538	-	-
Homo sapiens	P19099	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant mature form of C-terminally His6-tagged CYP11B1 from Escherichia coli strain JM109 by nickel affinity and cation exchange chromatography to homogeneity	Homo sapiens
recombinant mature form of C-terminally His6-tagged CYP11B2 from Escherichia coli strain JM109 by nickel affinity and cation exchange chromatography to homogeneity	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
11-deoxycorticosterone + reduced adrenodoxin + O2	-	Homo sapiens	corticosterone + oxidized adrenodoxin + H2O	-	?
11-deoxycorticosterone + reduced adrenodoxin + O2	i.e. 21-hydroxyprogesterone	Homo sapiens	corticosterone + oxidized adrenodoxin + H2O	-	?
11-deoxycortisol + reduced adrenodoxin + O2	-	Homo sapiens	cortisol + oxidized adrenodoxin + H2O	-	?
androstendione + reduced adrenodoxin + O2	low activity with isozyme CYP11B1	Homo sapiens	? + oxidized adrenodoxin + H2O	-	?
androstendione + reduced adrenodoxin + O2	low activity with isozyme CYP11B2	Homo sapiens	? + oxidized adrenodoxin + H2O	-	?
corticosterone + reduced adrenodoxin + O2	-	Homo sapiens	18-hydroxycorticosterone + oxidized adrenodoxin + H2O	-	?
additional information	aldosterone synthase shows high 11beta-hydroxylase activity toward both gluco- and mineralocorticoid formation, and additional 18-oxidase activity, substrate specificties of isozymes CYP11B2 and CYP11B1, overview. No oxidation activity of CYP11B2 with 18-hydroxycorticosterone, 11-dehydrocorticosterone, 21-hydroxypregnenolone, and poor activity with cortisol	Homo sapiens	?	-	?
additional information	substrate specificties of isozymes CYP11B2 and CYP11B1, overview. No or poor activity of CYP11B1 with cortisol, 18-hydroxycorticosterone, 11-dehydrocorticosterone, 21-hydroxypregnenolone, and 11beta-hydroxyprogesterone	Homo sapiens	?	-	?
progesterone + reduced adrenodoxin + O2	low activity with isozyme CYP11B1	Homo sapiens	11beta-hydroxyprogesterone + oxidized adrenodoxin + H2O	-	?
progesterone + reduced adrenodoxin + O2	low activity with isozyme CYP11B2	Homo sapiens	11beta-hydroxyprogesterone + oxidized adrenodoxin + H2O	-	?
testosterone + reduced adrenodoxin + O2	low activity with isozyme CYP11B1	Homo sapiens	11-hydroxytestosterone + oxidized adrenodoxin + H2O	-	?
testosterone + reduced adrenodoxin + O2	low activity with isozyme CYP11B2	Homo sapiens	11-hydroxytestosterone + oxidized adrenodoxin + H2O	-	?

Synonyms

Synonyms	Comment	Organism
Aldosterone synthase	-	Homo sapiens
CYP11B1	-	Homo sapiens
CYP11B2	-	Homo sapiens
cytochrome P450 11B1	-	Homo sapiens
cytochrome P450 11B2	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
adrenodoxin	-	Homo sapiens

General Information

General Information	Comment	Organism
evolution	although the active site pocket is lined by identical residues between CYP11B isoforms, most of the divergent residues that confer additional 18-oxidase activity of aldosterone synthase are located in the I-helix, vicinity of the O2 activation path and loops around the H-helix, affecting an egress channel closure required for retaining intermediates in the active site	Homo sapiens
additional information	a hydrophobic cavity with specific features is associated with corticosteroid recognition, substrate binding mode, overview	Homo sapiens
physiological function	aldosterone synthase, CYP11B2, is the sole enzyme responsible for the production of aldosterone in humans	Homo sapiens