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Literature summary extracted from

  • Bednarski, B.; Andreesen, J.R.; Pich, A.
    In vitro processing of the proproteins GrdE of protein B of glycine reductase and PrdA of D-proline reductase from Clostridium sticklandii: formation of a pyruvoyl group from a cysteine residue (2001), Eur. J. Biochem., 268, 3538-3544.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.21.4.1 cloning of the genes encoding D-proline reductase, 4.8 kb EcoRI fragment containing the genes prdA and prdB isolated and sequenced, prdAcodes for a 68 kDa protein, posttranslationally cleaved to the 45 kDa and 23 kDa subunits, prdB encodes the 26 kDa subunit Acetoanaerobium sticklandii
1.21.4.1 D-proline reductase proproteins cloned and overexpressed in Escherichia coli XL2 blue and BL21 Acetoanaerobium sticklandii
1.21.4.2 of the pyruvoyl group-forming proproteins grdE, expression in Escherichia coli Acetoanaerobium sticklandii

Protein Variants

EC Number Protein Variants Comment Organism
1.21.4.2 C242A no cleavage on the N-terminal side of a cysteine Acetoanaerobium sticklandii
1.21.4.2 C242S cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme Acetoanaerobium sticklandii
1.21.4.2 C242T cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme Acetoanaerobium sticklandii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.21.4.1 23000
-
 10 * 23000 + 10 * 26000+ 10 * 45000, SDS-PAGE, mass spectrometry Acetoanaerobium sticklandii
1.21.4.1 45000
-
 10 * 23000 + 10 * 26000+ 10 * 45000, SDS-PAGE, mass spectrometry Acetoanaerobium sticklandii
1.21.4.2 21162
-
 selenoprotein B, alpha,beta, 1 * 21162 + 1 * 26295, mass spectroscopy, amino acid sequence Acetoanaerobium sticklandii
1.21.4.2 26295
-
 selenoprotein B, alpha,beta, 1 * 21162 + 1 * 26295, mass spectroscopy, amino acid sequence Acetoanaerobium sticklandii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O Acetoanaerobium sticklandii
-
 glycine + phosphate + thioredoxin
-
 ?
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O Acetoanaerobium sticklandii DSM 519
-
 glycine + phosphate + thioredoxin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.21.4.1 Acetoanaerobium sticklandii
-
 strain ATCC12662 identical with strain DSM 519 T
-
1.21.4.2 Acetoanaerobium sticklandii P26971 strain HF, DSM 519T from DSMZ
-
1.21.4.2 Acetoanaerobium sticklandii DSM 519 P26971 strain HF, DSM 519T from DSMZ
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.21.4.1
-
 Acetoanaerobium sticklandii
1.21.4.2 of pyruvoyl group-forming protein grdE Acetoanaerobium sticklandii

Reaction

EC Number Reaction Comment Organism Reaction ID
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin mechanism Acetoanaerobium sticklandii
a
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin cleavage mechanism for the pyruvoyl group dependent reductase starting from cysteine Acetoanaerobium sticklandii
a
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase) Acetoanaerobium sticklandii
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.21.4.1 D-proline + dithiothreitol Stickland reaction Acetoanaerobium sticklandii 5-amino pentanoic acid + ?
-
 ?
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O
-
 Acetoanaerobium sticklandii glycine + phosphate + thioredoxin
-
 ?
1.21.4.2 acetyl phosphate + NH3 + thioredoxin disulfide + H2O
-
 Acetoanaerobium sticklandii DSM 519 glycine + phosphate + thioredoxin
-
 ?

Subunits

EC Number Subunits Comment Organism
1.21.4.1 decamer 10 * 23000 + 10 * 26000+ 10 * 45000, SDS-PAGE, mass spectrometry Acetoanaerobium sticklandii
1.21.4.2 dimer selenoprotein B, alpha,beta, 1 * 21162 + 1 * 26295, mass spectroscopy, amino acid sequence Acetoanaerobium sticklandii