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Literature summary extracted from

  • Verdoucq, L.; Moriniere, J.; Bevan, D.R.; Esen, A.; Vasella, A.; Henrissat, B.; Czjze, M.
    Structural determinants of substrate specificity in family 1 beta-glucosidases: novel insights from the crystal structure of sorghum dhurrinase-1, a plant beta-glucosidase with strict specificity, in complex with its natural substrate (2004), J. Biol. Chem., 279, 31796-31803.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 overexpression of Dhr1 and mutant E189D in Escherichia coli as His-tagged proteins Sorghum bicolor

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.21 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
3.2.1.21 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

EC Number Protein Variants Comment Organism
3.2.1.21 E189D substrate binding structure analysis from crystal structure Sorghum bicolor
3.2.1.21 E191D substrate binding structure analysis from crystal structure Zea mays
3.2.1.21 E191D/F198V inactive mutant, F198 substitution causes a rearrangement of residues F205, F466, and E464 involved in aglycone binding Zea mays

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.21 cyclic glucotetrazole mimics the 4H3 transition state intermediate Sorghum bicolor
3.2.1.21 cyclic glucotetrazole mimics the 4H3 transition state intermediate Zea mays

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.21 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
-
 ?
3.2.1.21 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

EC Number Organism UniProt Comment Textmining
3.2.1.21 Sorghum bicolor
-
 isozyme Dhr1
-
3.2.1.21 Zea mays P49235
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.21 recombinant His-tagged Dhr1 and mutant E189D from Escherichia coli by nickel affinity chromatography Sorghum bicolor

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.21 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
a
3.2.1.21 celloheptaose + 6 H2O = 7 beta-D-glucose substrate binding mode and structure Zea mays
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 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
-
 ?
3.2.1.21 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
-
 ?
3.2.1.21 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
-
 ?
3.2.1.21 additional information Glu1 shows broad substrate specificity in vitro Zea mays ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.21 Dhr1
-
 Sorghum bicolor
3.2.1.21 dhurrinase-1
-
 Sorghum bicolor
3.2.1.21 Glu1
-
 Zea mays