Cloned (Comment) | Organism |
---|---|
overexpression of Dhr1 and mutant E189D in Escherichia coli as His-tagged proteins | Sorghum bicolor |
Crystallization (Comment) | Organism |
---|---|
purified recombinant Glu1 mutant E191D in complex with cyclic inhibitor glucoetrazole, the crystallization solution is miced with 0.005 ml glycerol and 0.003 ml of inhibitor solution, X-ray diffraction structure determination and analysis at 1.95 A resolution | Zea mays |
wild-type Dhr1, and Dhr1 mutant E189D in complex with dhurrin, hanging drop vapour diffusion method, 0.002 ml of 8 mg/ml protein in 20 mM Tris, pH 7.0, mixed with an equal volume of reservoir solution containing 0.1 M HEPES, pH 7.5, 2.0 M ammonium sulfate, and 50 mM NaCl, X-ray diffraction structure determination and analysis at 1.9-2.0 A resolution | Sorghum bicolor |
Protein Variants | Comment | Organism |
---|---|---|
E189D | substrate binding structure analysis from crystal structure | Sorghum bicolor |
E191D | substrate binding structure analysis from crystal structure | Zea mays |
E191D/F198V | inactive mutant, F198 substitution causes a rearrangement of residues F205, F466, and E464 involved in aglycone binding | Zea mays |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
cyclic glucotetrazole | mimics the 4H3 transition state intermediate | Sorghum bicolor | |
cyclic glucotetrazole | mimics the 4H3 transition state intermediate | Zea mays |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | Zea mays | - |
2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose | - |
? | |
dhurrin + H2O | Sorghum bicolor | i.e. 4-hydroxy-(S)-mandelonitrile-beta-D-glucoside, Dhr1 is strictly specific for this substrate | 4-hydroxy-(S)-mandelonitrile + beta-D-glucose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sorghum bicolor | - |
isozyme Dhr1 | - |
Zea mays | P49235 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged Dhr1 and mutant E189D from Escherichia coli by nickel affinity chromatography | Sorghum bicolor |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
celloheptaose + 6 H2O = 7 beta-D-glucose | structural basis for substrate specificity, residues N259, F261, and S462 are important, substrate binding mode and structure | Sorghum bicolor | |
celloheptaose + 6 H2O = 7 beta-D-glucose | substrate binding mode and structure | Zea mays |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | - |
Zea mays | 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose | - |
? | |
2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one + H2O | i.e. DIMBOA-glucoside | Zea mays | 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one + beta-D-glucose | - |
? | |
dhurrin + H2O | i.e. 4-hydroxy-(S)-mandelonitrile-beta-D-glucoside, Dhr1 is strictly specific for this substrate | Sorghum bicolor | 4-hydroxy-(S)-mandelonitrile + beta-D-glucose | - |
? | |
additional information | Glu1 shows broad substrate specificity in vitro | Zea mays | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Dhr1 | - |
Sorghum bicolor |
dhurrinase-1 | - |
Sorghum bicolor |
Glu1 | - |
Zea mays |