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Literature summary for 3.6.5.2 extracted from

  • Kosami, K.; Ohki, I.; Nagano, M.; Furuita, K.; Sugiki, T.; Kawano, Y.; Kawasaki, T.; Fujiwara, T.; Nakagawa, A.; Shimamoto, K.; Kojima, C.
    The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase (2014), J. Biol. Chem., 289, 28569-28578.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
OsRac1, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally GST-tagged wild-type and mutant enzymes in Escherichia coli Rosetta (DE3), transient coexpression of OsRac1 andthe NADPH oxidase OsRbohB in Nicotiana benthamiana leaves enhancing ROS production Oryza sativa

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant GST-tagged OsRac1 complexed with non-hydrolyzable GTP analogue guanosine 5'-(beta,gamma-imido)triphosphate, sitting-drop vapor-diffusion method, mixing of 0.007 ml of 4 mg/ml protein with 0.007 ml of reservoir solution containing 100mM MES, pH 6.0, and 10-30% PEG 6000, 25% v/v glycerol as a cryoprotectant, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement Oryza sativa

Protein Variants

Protein Variants Comment Organism
C32S/Q68L design OsRac1 mutants that display reduced binding to the NADPH oxidase OsRbohB. Tyr39 and Asp45 substitutions suppress ROS production in rice cells, these residues are critical for interaction with and activation of NADPH oxidase OsRbohB Oryza sativa
G19V the constitutively activated mutant of OsRac1 increases resistance to rice bacterial blight disease and subsequent cell death Oryza sativa
G19V/D38A site-directed mutagenesis, the Switch I mutant shows unaltered binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/D45A site-directed mutagenesis, the Switch I mutant shows markedly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/F35A site-directed mutagenesis, the Switch I mutant shows markedly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/F44A site-directed mutagenesis, the Switch I mutant shows moderately reduced binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/I40A site-directed mutagenesis, the Switch I mutant shows slightly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/P36A site-directed mutagenesis, the Switch I mutant shows almost unaltered binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/P41A site-directed mutagenesis, the Switch I mutant shows slightly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/T37A site-directed mutagenesis, the Switch I mutant shows slightly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/T42A site-directed mutagenesis, the Switch I mutant shows markedly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/V43A site-directed mutagenesis, the Switch I mutant shows moderately reduced binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
G19V/Y39A site-directed mutagenesis, the Switch I mutant shows markedly attenuated binding with OsRbohB(138-313) compared to the wild-type enzyme Oryza sativa
T24N a dominant-negative mutant of OsRac1 that decrease the resistance reaction to rice bacterial blight disease and subsequent cell death Oryza sativa

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Oryza sativa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GTP + H2O Oryza sativa
-
GDP + phosphate
-
?
additional information Oryza sativa interaction site of HsRac1 with p67phox, an essential component of the NADPH oxidase complex ?
-
?

Organism

Organism UniProt Comment Textmining
Oryza sativa Q9SSX0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally GST-tagged wild-type and mutant enzymes from Escherichia coli Rosetta (DE3) by glutathione affinity chromatography Oryza sativa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GTP + H2O
-
Oryza sativa GDP + phosphate
-
?
additional information interaction site of HsRac1 with p67phox, an essential component of the NADPH oxidase complex Oryza sativa ?
-
?

Synonyms

Synonyms Comment Organism
OsRac1
-
Oryza sativa
Rho-type small GTPase
-
Oryza sativa
small GTPase
-
Oryza sativa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Oryza sativa

General Information

General Information Comment Organism
metabolism Rac/Rop proteins are Rho-type small GTPases that act as molecular switches in plants. They are key components in many major plant signaling pathways, such as innate immunity, pollen tube growth, and root hair formation Oryza sativa
additional information the solvent accessibility values of all residues located in Switch I are for Phe35, Tyr39, Thr42, Val43, Phe44, and Asp45 16, 100, 55, 54, 54, and 69%, respectively Oryza sativa
physiological function Rac/Rop protein OsRac1 plays an important role in regulating the production of reactive oxygen species by the NADPH oxidase OsRbohB during innate immunity. Direct OsRac1-OsRbohB interactions activate NADPH oxidase in plants Oryza sativa