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Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.14.16 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)

EC Number Crystallization (Comment) Organism
1.14.14.16 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.14.16 additional information
-
 additional information Pre-steady-state and steady-state binding kinetics with 17alpha-hydroxyprogesterone, stopped-flow spectroscopic measurements, overview Bos taurus
1.14.14.16 0.0019
-
 17alpha-hydroxyprogesterone pH 7.4, 37°C, wild-type CYP21 Bos taurus
1.14.14.16 0.0024
-
 17alpha-hydroxyprogesterone pH 7.4, 37°C, CYP21 mutant T241R/L442A Bos taurus

Metals/Ions

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.14.16 17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2 Bos taurus
-
 11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2 Homo sapiens
-
 11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 progesterone + [reduced NADPH-P450 reductase] + O2 Bos taurus
-
 11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 progesterone + [reduced NADPH-P450 reductase] + O2 Homo sapiens
-
 11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
 ?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.16 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.16 17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
-
 Bos taurus 11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 17alpha-hydroxyprogesterone + [reduced NADPH-P450 reductase] + O2
-
 Homo sapiens 11-deoxycortisol + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 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
-
 ?
1.14.14.16 progesterone + [reduced NADPH-P450 reductase] + O2
-
 Bos taurus 11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
 ?
1.14.14.16 progesterone + [reduced NADPH-P450 reductase] + O2
-
 Homo sapiens 11-deoxycorticosterone + [oxidized NADPH-P450 reductase] + H2O
-
 ?

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.16 37
-
 assay at Bos taurus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.14.16 0.22
-
 17alpha-hydroxyprogesterone pH 7.4, 37°C, CYP21 wild-type and mutant T241R/L442A Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.16 7.4
-
 assay at Bos taurus

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.16 heme
-
 Bos taurus

General Information

EC Number General Information Comment Organism
1.14.14.16 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
1.14.14.16 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