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Literature summary extracted from

  • Mahanta, N.; Liu, A.; Dong, S.; Nair, S.; Mitchell, D.
    Enzymatic reconstitution of ribosomal peptide backbone thioamidation (2018), Proc. Natl. Acad. Sci. USA, 115, 3030-3035 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.2.2.1 additional information TfuA may allosterically activate the enzyme or assist in the delivery of sulfide to the substrate Methanocaldococcus jannaschii
6.2.2.1 additional information TfuA may allosterically activate the enzyme or assist in the delivery of sulfide to the substrate Methanosarcina acetivorans
6.2.2.1 additional information TfuA may allosterically activate the enzyme or assist in the delivery of sulfide to the substrate Methanopyrus kandleri

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.2.2.1 expressed in Escherichia coli BL21-RIPL (DE3) cells Methanocaldococcus jannaschii
6.2.2.1 expressed in Escherichia coli BL21-RIPL (DE3) cells Methanosarcina acetivorans
6.2.2.1 expressed in Escherichia coli BL21-RIPL (DE3) cells Methanopyrus kandleri

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.2.2.1 enzyme in complex with ACP-Mg2+, hanging drop vapor diffusion method, using 0.02 M sodium formate, 0.02 M ammonium acetate, 0.02 M sodium citrate (tribasic), 0.02 M sodium potassium tartrate, 0.02 M sodium oxamate, 0.05 M HEPES pH 7.5, 0.05 M MOPS pH 7.5, 20% (v/v) glycerol, and 10% (w/v) PEG 4000 Methanopyrus kandleri

Protein Variants

EC Number Protein Variants Comment Organism
6.2.2.1 D40A the mutant shows reduced activity compared to the wild type enzyme Methanopyrus kandleri
6.2.2.1 D48A the mutant shows no activity Methanosarcina acetivorans
6.2.2.1 E156A the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 E164A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 E167A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 E201A the mutant shows no activity Methanosarcina acetivorans
6.2.2.1 E251A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 E78A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 E81A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 E87A the mutant shows no activity Methanosarcina acetivorans
6.2.2.1 E90A the mutant shows no activity Methanosarcina acetivorans
6.2.2.1 K408* the mutant shows wild type activity Methanosarcina acetivorans
6.2.2.1 K419* the mutant shows wild type activity Methanosarcina acetivorans
6.2.2.1 K64A the mutant shows reduced activity compared to the wild type enzyme Methanopyrus kandleri
6.2.2.1 M367* the mutant shows reduced activity compared to the wild type enzyme Methanopyrus kandleri
6.2.2.1 P16A the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 P372* the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 Q160A the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 Q254A the mutant shows no activity Methanopyrus kandleri
6.2.2.1 Q70A the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 R12A the mutant shows wild type activity Methanopyrus kandleri
6.2.2.1 R247A the mutant shows reduced activity compared to the wild type enzyme Methanopyrus kandleri
6.2.2.1 R40A the mutant shows wild type activity Methanosarcina acetivorans
6.2.2.1 R414* the mutant shows wild type activity Methanosarcina acetivorans
6.2.2.1 S74A the mutant shows reduced activity compared to the wild type enzyme Methanopyrus kandleri
6.2.2.1 W421* the mutant shows wild type activity Methanosarcina acetivorans

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.2.2.1 ALGFFGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 ALGFFGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 ALGFFGFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RAGFFGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RAGFFGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RAGFFGFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLAFFGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLAFFGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RLAFFGFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGAFGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGAFGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RLGAFGFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFAGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFAGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFAGFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFAFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFAFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFAFDLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGADLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGADLQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGADLQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGFALQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGFALQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGFALQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGFDAQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGFDAQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGFDAQD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGFDLAD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGFDLAD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGFDLAD
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGFDLQA
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGFDLQA
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGFDLQA
-
 Methanosarcina acetivorans
6.2.2.1 RLGFFGFDLQD
-
 Methanocaldococcus jannaschii
6.2.2.1 RLGFFGFDLQD
-
 Methanopyrus kandleri
6.2.2.1 RLGFFGFDLQD
-
 Methanosarcina acetivorans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.2.2.1 Mg2+ 20 mM used in assay conditions Methanocaldococcus jannaschii
6.2.2.1 Mg2+ 20 mM used in assay conditions Methanosarcina acetivorans
6.2.2.1 Mg2+ 20 mM used in assay conditions Methanopyrus kandleri

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465 Methanocaldococcus jannaschii
-
 ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465 Methanosarcina acetivorans
-
 ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465 Methanopyrus kandleri
-
 ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.2.2.1 Methanocaldococcus jannaschii
-
-
-
6.2.2.1 Methanopyrus kandleri Q8TZ25
-
-
6.2.2.1 Methanosarcina acetivorans
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.2.1 amylose resin column chromatography and Ni-NTA column chromatography Methanocaldococcus jannaschii
6.2.2.1 amylose resin column chromatography and Ni-NTA column chromatography Methanosarcina acetivorans
6.2.2.1 amylose resin column chromatography and Ni-NTA column chromatography Methanopyrus kandleri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.2.1 ATP + sulfide + RLGFFGFDLQD
-
 Methanocaldococcus jannaschii ADP + phosphate + RLGFF(thioglycine)FDLQD
-
 ?
6.2.2.1 ATP + sulfide + RLGFFGFDLQD
-
 Methanosarcina acetivorans ADP + phosphate + RLGFF(thioglycine)FDLQD
-
 ?
6.2.2.1 ATP + sulfide + RLGFFGFDLQD
-
 Methanopyrus kandleri ADP + phosphate + RLGFF(thioglycine)FDLQD
-
 ?
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465
-
 Methanocaldococcus jannaschii ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465
-
 Methanosarcina acetivorans ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?
6.2.2.1 ATP + sulfide + [methyl-coenzyme M reductase alpha-subunit McrA]-glycine465
-
 Methanopyrus kandleri ADP + phosphate + [methyl-coenzyme M reductase alpha-subunit McrA]-thioglycine465
-
 ?

Subunits

EC Number Subunits Comment Organism
6.2.2.1 monomer 1 * about 80000, SDS-PAGE Methanocaldococcus jannaschii
6.2.2.1 monomer 1 * about 80000, SDS-PAGE Methanosarcina acetivorans
6.2.2.1 monomer 1 * about 80000, SDS-PAGE Methanopyrus kandleri

Synonyms

EC Number Synonyms Comment Organism
6.2.2.1 ycaO
-
 Methanocaldococcus jannaschii
6.2.2.1 ycaO
-
 Methanosarcina acetivorans
6.2.2.1 ycaO
-
 Methanopyrus kandleri

Cofactor

EC Number Cofactor Comment Organism Structure
6.2.2.1 ATP
-
 Methanocaldococcus jannaschii
6.2.2.1 ATP
-
 Methanosarcina acetivorans
6.2.2.1 ATP
-
 Methanopyrus kandleri

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.2.2.1 0.0007
-
 RLGFFGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.00077
-
 ALGFFGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0009
-
 RLGFFGADLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0022
-
 RLGFFGFDLQA at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.008
-
 RLGFAGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0083
-
 RLGFFGFALQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0096
-
 RLGFFAFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0102
-
 RLGFFGFDAQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0103
-
 RLAFFGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0169
-
 RLGFFGFDLAD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.02
-
 RLGAFGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans
6.2.2.1 0.0201
-
 RAGFFGFDLQD at pH 7.5 and 25°C Methanosarcina acetivorans