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 (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 | 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Oryza sativa |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Oryza sativa | Q9SSX0 | - |
- |
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 | 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 | Comment | Organism |
---|---|---|
OsRac1 | - |
Oryza sativa |
Rho-type small GTPase | - |
Oryza sativa |
small GTPase | - |
Oryza sativa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Oryza sativa |
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 |