| acceptable | ECOLI:DYR | 2016-04-04 09:47:49 CDT | GO:0004146 dihydrofolate reductase activity (F) | PMID:26147643 | ECO:0000315 mutant phenotype evidence used in manual assertion | Typically, dihydrofolate reductase (DHFR) catalyzes the NADPH-dependent reduction of dihydrofolate (DHF) to tetrahydrofolate (THF). DHFR acts in a catalytic cycle of 5 different intermediates. In the mutant (by changing Leu 28 to Phe), the dissociation rate of the enzyme:substrate complex increases by 10-20 fold. In addition, the rates of the exchange of the excited closed states are very different in the mutants, despite the similarities found in the ground states of the mutant and wild-types. There are 2 pathways, intrinsic and allosteric pathways that this enzyme can take depending on the environmental conditions.
| challenge |
| unacceptable | 9CAUD:A0A0K2D000 | 2016-04-04 09:42:34 CDT | GO:0004146 dihydrofolate reductase activity (F) | PMID:26147643 | ECO:0000250 sequence similarity evidence used in manual assertion | Transfer annotation from gene page ECOLI:DYR.
With an E value of 2e-35, there is strong support for sequence similarity (accession #: 5CC9_A).
| challenge |
| unacceptable | LACCA:DYR | 2016-03-27 12:20:54 CDT | GO:0050661 NADP binding (F) | PMID:6815179 | ECO:0000314 direct assay evidence used in manual assertion | Using crystallography, scientists determined that when NADPH binds to dihydrofolate reductase, the enzyme:substrate complex geometry stands out. Nicotinamide interacts with 3 oxygen atoms and 3 hydrogen atoms in this very polar environment.
| challenge |
| unacceptable | 9CAUD:A0A0K2D000 | 2016-04-04 07:03:18 CDT | GO:0050661 NADP binding (F) | PMID:6815179 | ECO:0000250 sequence similarity evidence used in manual assertion | Transfer annotation from gene page LACCA:DYR.
Strong sequence similarity with an E value of 2E-34.
Accession #: 4DFR_A
| challenge |
| unacceptable | THEMA:DYR | 2016-04-04 10:53:39 CDT | GO:0004146 dihydrofolate reductase activity (F) | PMID:10731419 | ECO:0000314 direct assay evidence used in manual assertion | The enzyme does not show much structural changes when bound to its coenzyme NADPH and its inhibitor methotrexate. The enzyme also forms a homodimer, which contributes to its extreme stability.
| challenge |
| unacceptable | 9CAUD:A0A0K2D000 | 2016-04-04 10:59:32 CDT | GO:0004146 dihydrofolate reductase activity (F) | PMID:10731419 | ECO:0000250 sequence similarity evidence used in manual assertion | With an E value of 1e-14, there is strong support for sequence similarity (accession #: WP_010865375.1).
| challenge |
| unacceptable | THEMA:DYR | 2016-04-04 11:14:59 CDT | GO:0004146 dihydrofolate reductase activity (F) | PMID:10413491 | ECO:0000314 direct assay evidence used in manual assertion | The enzymes were not found to exist as monomers in equilibrium or while unfolding. This suggests that the protein is extremely stable throughout experimental temperatures from 5-70 degrees Celsius with a maximum stability at about 35 degrees C.
| challenge |
| acceptable | BACSU:MCSA | 2016-04-07 08:07:06 CDT | GO:0006508 proteolysis (P) | PMID:11179229 | ECO:0000315 mutant phenotype evidence used in manual assertion | McsA play a role in regulating transcription of CtsR-dependent heat shock genes. Figure 5- In the mcsA mutants, CtsR-dependent heat shock proteins under non-stressed conditions were increased when compared with the wild-type.
Figure 6B- In nonstressed McsA mutant cells, the CtsR level was lower than it was in the wild type. After heat shock, CtsR levels in the mutant decreased suggestin that CtsR is less stable without McsA. The half-life of CtsR was also lower in cells lacking McsA
| challenge |
| unacceptable | BACSU:MCSA | 2016-04-07 07:58:58 CDT | GO:0006508 proteolysis (P) | PMID:6508 | ECO:0000314 direct assay evidence used in manual assertion | Through gel mobility shift experiments (Figure 6), it was shown that addition of McsA does not influence he CtsR DNA binding activity while excess (12-fold) McsA could promote CtsR-dependent DNA binding.
| challenge |
| unacceptable | BACSU:MCSB | 2016-04-07 08:18:55 CDT | GO:0006508 proteolysis (P) | PMID:11179229 | ECO:0000315 mutant phenotype evidence used in manual assertion | McsB play a role in regulating transcription of CtsR-dependent heat shock genes. Figure 5- In the mcsB mutants, CtsR-dependent heat shock proteins under non-stressed conditions were increased when compared with the wild-type. McsB influences the CtsR-mediated repression negatively. Figure 6B- The half-life of the CtsR repressor was increased in McsB mutants, suggesting that McsB destabilizes CtsR.
| challenge |
| unacceptable | BACSU:MCSB | 2016-04-07 08:18:21 CDT | GO:0006508 proteolysis (P) | PMID:11179229 | ECO:0000314 direct assay evidence used in manual assertion | Through gel mobility shift experiments (Figure 6, lanes 8-10), it was shown that addition of McsB to CtsR led to a decrease in the amount of DNA retarded by CtsR. This suests that McsB can remove the DNA binding capacity of CtsR.
| challenge |
