EC Number   |
Protein Variants |
Reference |
|---|
   7.1.1.10 | more |
in DELTAndhD1 and DELTAndhD1/D2 mutants, the NDH-1L complex is missing but NDH-1M and NDH-1S are abundant in low-CO2 conditions. The DELTAndhD3 and DELTAndhD3/D4 mutants lack NDH-1S but exhibit wild-type levels of NDH-1M at low CO2. In both mutants, the carbon uptake is impaired and more severely in the double mutant that can grow at low CO2 only at elevated pH (pH 8.3 but not 7.5), when other mechanisms of carbon uptake are activated. Deletion of the small hydrophobic NdhL subunit is not essential for the assembly of NDH-1L and NDH-1M. This subunit might be important in supporting a specific conformation of the NDH-1 complexes because the importance of NdhL for the function in CO2 uptake is unambiguously demonstrated |
762110 |
| 7.1.1.10 | more |
preparation of NDH-Fd supercomplex: the purified NDH-1L complex is concentrated to 1 mg/ml and mixed with NdhS, NdhV, and Fd proteins. The molar ratio of NDH-1L:NdhS:NdhV:Fd is set to 1:4:6:6. The mixture is incubated for 1 h at 20°C and then concentrated to 12 mg/ml |
761986 |
| 7.1.1.10 | more |
reoxidation of P700 is much faster in DELTAcpcG2 than in the wild-type. Furthermore, the rereduction rate of P700+ was monitored in darkness after the illumination of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-treated cells with FR. The rereduction of P700+ is much slower in DELTAcpcG2 than in the wild-type, providing evidence of the scarcity of CET from reduced Fd via NDH-1 back to P700+ in darkness. Therefore, the slow growth of DELTAcpcG2 under high-light conditions can be attributed to the low NDH-CET activity. Complex analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, immunoblot and coimmunoprecipitation |
762156 |
| 7.1.1.10 | more |
screening of a transposon-tagged library of Synechocystis sp. 6803 cells grown under high light for subunit identification. Generation of mutantsx02ssl0352,x02ndhB (M55), Ssl0352-YFP-His6, and NdhM-YFP-His6 (two NDH-1-mediated CET (NDH-CET))-defective mutants are tagged in gene ssl0352. Construction of a ssl0352 deletion mutant and another mutant with Ssl0352 fused to yellow fluorescent protein (YFP) with a His6-tag are constructed. Immunoblotting, mass spectrometry, and confocal microscopy analyses reveal that the Ssl0352 protein resides in the thylakoid membrane and associates with the NDH-1L and NDH-1M complexes. Deletion of the ssl0352 gene considerably impairs the NDH-CET activity and also retards cell growth under high light conditions, indicating that NdhS is essential for efficient operation of NDH-CET. The high light-dependent phenotype of the two mutants results from defective NDH-CET. But the assembly of the NDH-1L and NDH-1M complexes and their content in the cells are not affected in the mutant |
761445 |
| 7.1.1.10 | more |
several strains of Thermosynechococcus elongatus are constructed by adding a His-tag to different subunits of NDH-1. Two strains with His-tag on CupA and NdhL are successfully used to isolate NDH-1 complexes by one-step Ni2+ column chromatography, revealing the presence of three complexes with molecular masses of about 450, 300 and 190 kDa, which are identified by MS to be NDH-1L, NDH-1M and NDH-1S respectively. A larger complex of about 490 kDa is also isolated from the NdhL-His strain. This complex, designated NDH-1MS, is composed of NDH-1M and NDH-1S (EC 7.1.1.11). Selection of the subunit for His tagging is crucial for isolation of NDH-1 complexes |
760518 |
| 7.1.1.10 | more |
the NDH-1L complex, missing from the DELTANdhD1/D2 mutant, is a prerequisite for photoheterotrophic growth and thus apparently involved in cellular respiration. The amount of NDH-1M and the rate of P700+ rereduction in darkness in the DELTANdhD1/D2 mutant grown at low CO2 are similar to those in the wild-type, whereas in the M55 mutant (DELTANdhB), lacking both NDH-1L and NDH-1M, the rate of P700+ rereduction is very slow. The NDH-1S (small) complex, localized to the thylakoid membrane and composed of only NdhD3, NdhF3, CupA, and Sll1735, is strongly induced at low CO2 in the wild-type as well as in DELTANdhD1/D2 and M55. In contrast with the wild-type and DELTANdhD1/D2, which show normal CO2 uptake, M55 is unable to take up CO2 even when the NDH-1S complex is present. Conversely, the DELTANdhD3/D4 mutant, also unable to take up CO2, lacks NDH-1S but exhibits wild-type levels of NDH-1M at low CO2 |
762122 |
