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Literature summary extracted from

  • Porte, S.; Moeini, A.; Reche, I.; Shafqat, N.; Oppermann, U.; Farres, J.; Pares, X.
    Kinetic and structural evidence of the alkenal/one reductase specificity of human zeta-crystallin (2011), Cell. Mol. Life Sci., 68, 1065-1077.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.27
-
Homo sapiens
1.6.5.5 expressed in Escherichia coli as a His-tagged fusion protein Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.3.1.27 to 1.85 A resolution. The overall structure displays the typical medium-chain dehydrogenase fold with two domains: the catalytic domain, residues 1-128 and 271-329, and the coenzyme-binding domain, residues 129-270, which are separated by a deep cleft that binds the cofactor NADP+ and the substrate Homo sapiens
1.6.5.5 crystallographic structure of human zeta-crystallin is reported. Enzyme shows a tetrameric structure Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.3.1.27 Y53F substrate-pocket residue, large increase in Km values for all substrates and the cofactor, but mainly towards 3-buten-2-one and propenal with a 30fold increase Homo sapiens
1.3.1.27 Y59F substrate-pocket residue, almost the same kinetic parameter values as the wild-type enzyme for 2-alkenals Homo sapiens
1.6.5.5 T53F Tyr53Phe mutant displays a large increase in Km values for all substrates and the cofactor, but mainly towards 3-buten-2-one and propenal with a 30fold increase. For 3-penten-2-one, 3-nonen-2-one, 2-hexenal and 4-hydroxy-2-hexenal mutant also shows a decrease in kcat Homo sapiens
1.6.5.5 T59F Tyr59Phe mutant exhibits almost the same kinetic parameter values as the wild-type enzyme for 2-alkenals,while the Km is increased for 4-hydroxy-2-hexenal Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.27 0.005
-
NADPH wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.015
-
NADPH mutant Y59F, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.035
-
3-buten-2-one wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.06
-
NADPH mutant Y53F, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.13
-
4-hydroxy-2-hexenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.48
-
2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.55
-
4-hydroxy-2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 1.26
-
4-hydroxy-2-hexenal mutant Y59F, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 3.67
-
4-hydroxy-2-hexenal mutant Y53F, pH 7.5, temperature not specified in the publication Homo sapiens
1.6.5.5 0.005
-
NADPH pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.015
-
NADPH pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.035
-
3-buten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.05
-
3-nonen-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.06
-
NADPH pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.06
-
3-penten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.06
-
3-buten-2-one pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.08
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.13
-
4-hydroxy-2-hexenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.13
-
3-buten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.13
-
2-hexenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.482
-
2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.535
-
3-penten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.55
-
4-hydroxy-2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.86
-
propenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.94
-
3-nonen-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 1.26
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 1.45
-
propenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 2.3
-
propenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 5.45
-
2-hexenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 6.12
-
2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 8.6
-
2-pentenal pH 7, 25°C, wild-type Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.27 Homo sapiens Q08257
-
-
1.6.5.5 Homo sapiens Q08257
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.6.5.5 using Ni-NTA chromatography Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.27 2-nonenal + NADPH + H+
-
Homo sapiens nonanal + NADP+
-
ir
1.3.1.27 3-buten-2-one + NADPH + H+
-
Homo sapiens butan-2-one + NADP+
-
?
1.3.1.27 4-hydroxy-2-hexenal + NADPH + H+
-
Homo sapiens 4-hydroxyhexadecanal + NADP+
-
?
1.3.1.27 4-hydroxy-2-nonenal + NADPH + H+
-
Homo sapiens 4-hydroxynonanal + NADP+
-
?
1.3.1.27 additional information enzyme additionally catalyzes the reduction of quinones, reaction of EC 1.6.5.5. The enzyme shows a low level of activity with cinnamaldehyde and no activity with 2-cyclohexen-1-one and 15-ketoprostaglandin E2. No activity is detected with nonanal and 2-nonene Homo sapiens ?
-
?
1.6.5.5 2-hexenal + NADPH
-
Homo sapiens hexanal + NADP+
-
?
1.6.5.5 2-nonenal + NADPH
-
Homo sapiens nonanal + NADP+
-
?
1.6.5.5 2-pentenal + NADPH
-
Homo sapiens pentanal + NADP+
-
?
1.6.5.5 3-buten-2-one + NADPH
-
Homo sapiens 2-butanone + NADP+
-
?
1.6.5.5 3-nonen-2-one + NADPH
-
Homo sapiens 2-nonanone + NADP+
-
?
1.6.5.5 3-penten-2-one + NADPH
-
Homo sapiens 2-pentanone + NADP+
-
?
1.6.5.5 4-hydroxy-2-hexenal + NADPH
-
Homo sapiens 4-hydroxy-hexanal + NADP+
-
?
1.6.5.5 4-hydroxy-2-nonenal + NADPH
-
Homo sapiens 4-hydroxy-nonanal + NADP+
-
?
1.6.5.5 propenal + NADPH
-
Homo sapiens propanal + NADP+
-
?

Subunits

EC Number Subunits Comment Organism
1.6.5.5 dimer gel filtration, 3 M NaCl Homo sapiens
1.6.5.5 homotetramer gel filtration, 0.3 M NaCl Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
1.3.1.27 zeta-Crystallin
-
Homo sapiens
1.6.5.5 zeta-Crystallin
-
Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.1.27 0.113
-
2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.167
-
3-buten-2-one wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 1.15
-
4-hydroxy-2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 1.33
-
4-hydroxy-2-hexenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.6.5.5 0.11
-
2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.16
-
3-buten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.2
-
3-penten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.2
-
2-pentenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.21
-
3-nonen-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.28
-
2-hexenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.28
-
3-buten-2-one pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.36
-
2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.97
-
3-buten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 1.15
-
4-hydroxy-2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 1.25
-
propenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 1.33
-
4-hydroxy-2-hexenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 1.66
-
propenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 2.16
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 2.66
-
3-penten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 3.11
-
3-nonen-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 3.67
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 39.74
-
propenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 73.27
-
2-hexenal pH 7, 25°C, mutant T53F Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.27 NADPH
-
Homo sapiens
1.6.5.5 NADPH
-
Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.3.1.27 0.02
-
4-hydroxy-2-hexenal mutant Y53F, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 0.23
-
2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 1.67
-
4-hydroxy-2-hexenal mutant Y59F, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 2.1
-
4-hydroxy-2-nonenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 4.8
-
3-buten-2-one wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.3.1.27 10.2
-
4-hydroxy-2-hexenal wild-type, pH 7.5, temperature not specified in the publication Homo sapiens
1.6.5.5 0.0016
-
2-hexenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.016
-
3-nonen-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.023
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.023
-
2-pentenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.025
-
3-penten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.051
-
2-hexenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.058
-
propenal pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.06
-
2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 0.13
-
3-buten-2-one pH 7, 25°C, mutant T53F Homo sapiens
1.6.5.5 0.23
-
2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.23
-
3-nonen-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 0.36
-
3-penten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 1.16
-
propenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 1.5
-
propenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 1.66
-
4-hydroxy-2-hexenal pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 2.1
-
4-hydroxy-2-nonenal pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 4.6
-
3-buten-2-one pH 7, 25°C, mutant T59F Homo sapiens
1.6.5.5 4.8
-
3-buten-2-one pH 7, 25°C, wild-type Homo sapiens
1.6.5.5 10.21
-
4-hydroxy-2-hexenal pH 7, 25°C, wild-type Homo sapiens