Literature summary for 1.14.14.16 extracted from

Zhao, B.; Lei, L.; Kagawa, N.; Sundaramoorthy, M.; Banerjee, S.; Nagy, L.D.; Guengerich, F.P.; Waterman, M.R.
Three-dimensional structure of steroid 21-hydroxylase (cytochrome P450 21A2) with two substrates reveals locations of disease-associated variants (2012), J. Biol. Chem., 287, 10613-10622.

Search for more literature for 1.14.14.16

Cloned(Commentary)

Cloned (Comment)	Organism
gene CYP21A2 or C3B21RA, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged wild-type enzyme and mutants in Escherichia coli	Bos taurus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged CYP21A2 mutant T241R/L442A in complex with substrate 17-hydroxyprogesterone, hanging drop vapor diffusion method, 0.2 mM protein in 50 mM potassium phosphate, pH 7.4, 20% glycerol, 0.1 mM DTT, 0.1 mM EDTA, 0.25% Cymal 5, and 50 mM NaCl, is mixed with 0.4 mM, containing 2% v/v C2H5OH, and 5-15% w/v PEG 3350, 0.5 M ammonium sulfate, and 0.1 M HEPES, pH 7.0, 20°C, few days, X-ray diffraction structure determination and analysis at 3.0 A resolution	Bos taurus

Crystallization (Comment)

Organism

purified recombinant His-tagged CYP21A2 mutant T241R/L442A in complex with substrate 17-hydroxyprogesterone, hanging drop vapor diffusion method, 0.2 mM protein in 50 mM potassium phosphate, pH 7.4, 20% glycerol, 0.1 mM DTT, 0.1 mM EDTA, 0.25% Cymal 5, and 50 mM NaCl, is mixed with 0.4 mM, containing 2% v/v C2H5OH, and 5-15% w/v PEG 3350, 0.5 M ammonium sulfate, and 0.1 M HEPES, pH 7.0, 20°C, few days, X-ray diffraction structure determination and analysis at 3.0 A resolution

Bos taurus

Protein Variants

Protein Variants	Comment	Organism
T241R	site-directed mutagenesis, the mutant shows improved solubility properties compared to the wild-type enzyme	Bos taurus
T241R/L442A	site-directed mutagenesis, the mutant shows greatly improved solubility properties compared to the wild-type enzyme	Bos taurus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Pre-steady-state and steady-state binding kinetics with 17alpha-hydroxyprogesterone, stopped-flow spectroscopic measurements, overview	Bos taurus
0.0019	-	17alpha-hydroxyprogesterone	pH 7.4, 37°C, wild-type CYP21	Bos taurus
0.0024	-	17alpha-hydroxyprogesterone	pH 7.4, 37°C, CYP21 mutant T241R/L442A	Bos taurus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	heme enzyme	Bos taurus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2	Bos taurus	-	11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O	-	?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2	Homo sapiens	-	11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O	-	?
progesterone + [reduced NADPH-P450 reductase] + O2	Bos taurus	-	11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O	-	?
progesterone + [reduced NADPH-P450 reductase] + O2	Homo sapiens	-	11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Bos taurus	P00191	gene CYP21A2 or C3B21RA	-
Homo sapiens	P08686	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type enzyme and mutants from Escherichia coli by nickel affinity and anion exchange chromatography, followed by gel filtration	Bos taurus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2	-	Bos taurus	11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O	-	?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2	-	Homo sapiens	11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O	-	?
17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2	two 17alpha-hydroxyprogesterone molecules are bound to the enzyme, the distal one being located at the entrance of the substrate access channel and the proximal one bound in the active site	Bos taurus	11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O	-	?
progesterone + [reduced NADPH-P450 reductase] + O2	-	Bos taurus	11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O	-	?
progesterone + [reduced NADPH-P450 reductase] + O2	-	Homo sapiens	11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O	-	?

Subunits

Subunits	Comment	Organism
More	structure-function analysis, overview	Bos taurus

Synonyms

Synonyms	Comment	Organism
C3B21RA protein	-	Bos taurus
CYP21A2	-	Bos taurus
CYP21A2	-	Homo sapiens
cytochrome p450 21A2	-	Bos taurus
cytochrome p450 21A2	-	Homo sapiens
Steroid 21-hydroxylase	-	Bos taurus
Steroid 21-hydroxylase	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Bos taurus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.22	-	17alpha-hydroxyprogesterone	pH 7.4, 37°C, CYP21 wild-type and mutant T241R/L442A	Bos taurus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Bos taurus

Cofactor

Cofactor	Comment	Organism	Structure
heme	-	Bos taurus

General Information

General Information	Comment	Organism
malfunction	steroid 21-hydroxylase deficiency accounts for about 95% of individuals with congenital adrenal hyperplasia, a common autosomal recessive metabolic disorder of adrenal steroidogenesis, mutations on CYP21A2 activity lead to impairment of the synthesis of cortisol and aldosterone and the excessive production of androgens	Homo sapiens
additional information	the key substrate recognition residues are not only around the heme but also along the substrate access channel, structure-function analysis, overview	Bos taurus