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Literature summary extracted from

  • Manszewski, T.; Singh, K.; Imiolczyk, B.; Jaskolski, M.
    An enzyme captured in two conformational states Crystal structure of S-adenosyl-L-homocysteine hydrolase from Bradyrhizobium elkanii (2015), Acta Crystallogr. Sect. D, 71, 2422-2432 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.13.2.1 expressed in Escherichia coli BL21 Star (DE3) cells Bradyrhizobium elkanii
3.13.2.1 expression in Escherichia coli BL21 Star (DE3) Bradyrhizobium elkanii
3.13.2.1 recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21 Star (DE3) Bradyrhizobium elkanii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.13.2.1 crystallization is conducted by vapour diffusion in hanging drops at 19°C. Crystal structure is solved at a resolution of 1.74 A, revealing a homotetramer in the asymmetric unit of space group P2(1)2(1)2 Bradyrhizobium elkanii
3.13.2.1 hanging drop vapor diffusion method, using 0.2 M ammonium acetate, 20%(w/v) polyethylene glycol 3350, pH 7.1 Bradyrhizobium elkanii
3.13.2.1 purified recombinant detagged enzyme, hanging drop vapour diffusion method, mixing of 12 mg/ml protein in 20 mM Tris, pH 8.0, 50 mM NaCl, with reservoir solution consisting of 0.2 M ammonium acetate, 20% w/v PEG 3350, pH 7.1, 18°C, X-ray diffraction structure determination and analysis at 1.74 A reoslution, molecular replacement using the atomic coordinates of chain A of SAHase from Brucella melitensis (PDB ID 3n58) as template, modelling Bradyrhizobium elkanii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.13.2.1 S-adenosyl-L-homocysteine + H2O Bradyrhizobium elkanii
-
 L-homocysteine + adenosine
-
 ?
3.13.2.1 S-adenosyl-L-homocysteine + H2O Bradyrhizobium elkanii the enzyme is involved in the enzymatic regulation of S-adenosyl-L-methionine (SAM)-dependent methylation reactions L-homocysteine + adenosine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.13.2.1 Bradyrhizobium elkanii A0A087WNH6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.13.2.1
-
 Bradyrhizobium elkanii
3.13.2.1 Ni-NTA column chromatography Bradyrhizobium elkanii
3.13.2.1 Ni-NTA column chromatography and gel filtration Bradyrhizobium elkanii
3.13.2.1 recombinant His6-tagged enzyme from Escherichia coli strain BL21 Star (DE3) by nickel affinity chromatography, tag cleavage by TEV protease, another step of nickel-affinity chromatography to remove the His-tag debris and His-tagged TEV protease, followed by gel filtration Bradyrhizobium elkanii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.13.2.1 S-adenosyl-L-homocysteine + H2O
-
 Bradyrhizobium elkanii L-homocysteine + adenosine
-
 ?
3.13.2.1 S-adenosyl-L-homocysteine + H2O the enzyme is involved in the enzymatic regulation of S-adenosyl-L-methionine (SAM)-dependent methylation reactions Bradyrhizobium elkanii L-homocysteine + adenosine
-
 ?

Subunits

EC Number Subunits Comment Organism
3.13.2.1 homotetramer 4 * 52000, SDS-PAGE Bradyrhizobium elkanii
3.13.2.1 homotetramer the protein crystallizes in its biologically relevant tetrameric form, a dimer of two tight dimers (AB and CD) Bradyrhizobium elkanii
3.13.2.1 More analysis of domain organization of substrate-binding domain, NAD+ cofactor-binding domain and dimerization domain in the subunits, quaternary interactions between the subunits, overview Bradyrhizobium elkanii
3.13.2.1 tetramer x * 52000, SDS-PAGE Bradyrhizobium elkanii
3.13.2.1 tetramer three subunits are in a closed conformation enforced by complex formation with the adenosine product of the enzymatic reaction. The fourth subunit is ligand-free and has an open conformation. Domain movement of the enzyme induced by the binding of its natural ligands. The centre of the tetramer is formed by the cofactor-binding domains, while the substrate-binding domains are located outside the core of the enzyme, where they are much more mobile than the rigid central part Bradyrhizobium elkanii

Synonyms

EC Number Synonyms Comment Organism
3.13.2.1 AHCY
-
 Bradyrhizobium elkanii
3.13.2.1 BeSAHase
-
 Bradyrhizobium elkanii
3.13.2.1 S-adenosyl-L-homocysteine hydrolase
-
 Bradyrhizobium elkanii
3.13.2.1 SAHase
-
 Bradyrhizobium elkanii

Cofactor

EC Number Cofactor Comment Organism Structure
3.13.2.1 NAD+
-
 Bradyrhizobium elkanii
3.13.2.1 NAD+ mode of binding and oxidation state, overview Bradyrhizobium elkanii

General Information

EC Number General Information Comment Organism
3.13.2.1 additional information the substrate-binding domain, built from amino acid residues Gly6-Val221 and Met397-Val426, has an alpha/beta-fold. The central parallel beta-sheet is built from seven beta-strands, structure overview. The C-terminal domain, formed by residues Leu427-Tyr473, has a helix-loop-helix-loop fold. Ligand-induced conformational change. Adenosine-binding site in the ligand-free subunit Bradyrhizobium elkanii
3.13.2.1 physiological function S-adenosyl-L-homocysteine hydrolase (SAHase) is involved in the enzymatic regulation of S-adenosyl-L-methionine (SAM)-dependent methylation reactions. After methyl-group transfer from SAM, S-adenosyl-L-homocysteine (SAH) is formed as a byproduct, which in turn is hydrolyzed to adenosine (Ado) and homocysteine (Hcy) by SAHase Bradyrhizobium elkanii
3.13.2.1 physiological function the enzyme is involved in the enzymatic regulation of S-adenosyl-L-methionine (SAM)-dependent methylation reactions Bradyrhizobium elkanii