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Literature summary extracted from

  • Ogata, K.; Yajima, Y.; Nakamura, S.; Kaneko, R.; Goto, M.; Ohshima, T.; Yoshimune, K.
    Inhibition of homoserine dehydrogenase by formation of a cysteine-NAD covalent complex (2018), Sci. Rep., 8, 5749 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.3 additional information the HSD from the hyperthermophilic archaeon Sulfolobus tokodaii (StHSD) is activated by reductive cleavage of the disulfide bond formed between cysteine residues (Cys304) in the C-terminal regions of the homodimer subunits Sulfurisphaera tokodaii

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.3 recombinant expression of the enzyme in Escherichia coli strain BL21(DE3) Sulfurisphaera tokodaii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.3 purified enzyme StHSD complexed with cysteine and NAD+, hanging drop vapour diffusion method, mixing of 0.002 ml of 5 mg/ml protein solution with 0.002 ml of reservoir solution containing 23% w/v PEG 3350, 0.2 M di-ammonium tartrate, 0.001 ml of 20 mM NAD+, and 0.001 ml of 100 mM cysteine, and equilibration against 0.1 ml of reservoir solution, 12°C, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement using the StHSD structure (PDB ID 4YDR) as an initial phasing model Sulfurisphaera tokodaii

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.3 L-cysteine competitive versus L-homoserine, uncompetitive versus cofactors NAD+ and NADP+. 95% inhibition at 10 mM. The feedback inhibition of StHSD by cysteine occurs through the formation of an enzyme-NAD-cysteine complex. Cysteine situates within the homoserine binding site, formation of a covalent bond between cysteine and the nicotinamide ring. Cysteine interacts with six residues (Gly156, Thr157, Tyr183, Glu185, Asp191, and Lys200) in the StHSD active site, binding structure analysis, overview Sulfurisphaera tokodaii
1.1.1.3 L-serine 14% inhibition at 10 mM Sulfurisphaera tokodaii
1.1.1.3 additional information tHSD is poorly inhibited by less than 5% by 10 mM L-methionine, L-isoleucine, or L-threonine, all of which are final products in the aspartate pathway, and by L-lysine Sulfurisphaera tokodaii
1.1.1.3 NADH acts as a competitive inhibitor of NAD+, product inhibition, non-competitive inhibition versus L-homoserine Sulfurisphaera tokodaii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.3 0.33
-
 NAD+ pH 8.0, 30°C Sulfurisphaera tokodaii
1.1.1.3 1.2
-
 NADP+ pH 8.0, 30°C Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.3 L-homoserine + NAD+ Sulfurisphaera tokodaii
-
 L-aspartate 4-semialdehyde + NADH + H+
-
 r
1.1.1.3 L-homoserine + NAD+ Sulfurisphaera tokodaii DSM 16993
-
 L-aspartate 4-semialdehyde + NADH + H+
-
 r
1.1.1.3 L-homoserine + NADP+ Sulfurisphaera tokodaii
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 r
1.1.1.3 L-homoserine + NADP+ Sulfurisphaera tokodaii DSM 16993
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.3 Sulfurisphaera tokodaii F9VNG5
-
-
1.1.1.3 Sulfurisphaera tokodaii DSM 16993 F9VNG5
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
1.1.1.3 proteolytic modification the HSD from the hyperthermophilic archaeon Sulfolobus tokodaii (StHSD) is activated by reductive cleavage of the disulfide bond formed between cysteine residues (Cys304) in the C-terminal regions of the homodimer subunits Sulfurisphaera tokodaii

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.3 recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange chromatography, dialysis, and ultrafiltration Sulfurisphaera tokodaii

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.3 L-homoserine + NAD(P)+ = L-aspartate 4-semialdehyde + NAD(P)H + H+ L-homoserine oxidation by StHSD proceeds through a sequentially ordered mechanism in which NAD+ binds to the free form of the enzyme, after which L-homoserine binds to the enzyme-NAD+ complex Sulfurisphaera tokodaii
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.3 L-homoserine + NAD+
-
 Sulfurisphaera tokodaii L-aspartate 4-semialdehyde + NADH + H+
-
 r
1.1.1.3 L-homoserine + NAD+
-
 Sulfurisphaera tokodaii DSM 16993 L-aspartate 4-semialdehyde + NADH + H+
-
 r
1.1.1.3 L-homoserine + NADP+
-
 Sulfurisphaera tokodaii L-aspartate 4-semialdehyde + NADPH + H+
-
 r
1.1.1.3 L-homoserine + NADP+
-
 Sulfurisphaera tokodaii DSM 16993 L-aspartate 4-semialdehyde + NADPH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.3 dimer StHSD is composed of a nucleotide-binding region (residues 1-130 and 285-304), a dimerization region (residues 131-145 and 256-284), and a catalytic region (residues 146-255). Presence of a disulfide bond formed between two cysteine residues (position 304) in the C-terminal regions of the two subunits Sulfurisphaera tokodaii

Synonyms

EC Number Synonyms Comment Organism
1.1.1.3 hom
-
 Sulfurisphaera tokodaii
1.1.1.3 HSD
-
 Sulfurisphaera tokodaii
1.1.1.3 StHSD
-
 Sulfurisphaera tokodaii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.3 30
-
 assay at Sulfurisphaera tokodaii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.3 8
-
 assay at Sulfurisphaera tokodaii

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.3 NAD+ binding structure analysis, overview Sulfurisphaera tokodaii
1.1.1.3 NADH
-
 Sulfurisphaera tokodaii
1.1.1.3 NADP+ StHSD also exhibits activity with NADP+, but with much lower efficiency compared to NAD+ Sulfurisphaera tokodaii
1.1.1.3 NADPH
-
 Sulfurisphaera tokodaii

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.1.1.3 0.011
-
 versus L-homoserine, pH 8.0, 30°C Sulfurisphaera tokodaii L-cysteine
1.1.1.3 0.55
-
 versus NAD+, pH 8.0, 30°C Sulfurisphaera tokodaii L-cysteine
1.1.1.3 1.2
-
 versus NADP+, pH 8.0, 30°C Sulfurisphaera tokodaii L-cysteine

General Information

EC Number General Information Comment Organism
1.1.1.3 physiological function homoserine dehydrogenase (HSD) is an important regulatory enzyme in the aspartate pathway, which mediates synthesis of methionine, threonine and isoleucine from aspartate Sulfurisphaera tokodaii