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| Content Provider | PubMed Central |
|---|---|
| Author | Zhang, Pengpeng Natalia, Battchikova Paakkarinen, Virpi Katoh, Hirokazu Iwai, Masako Ikeuchi, Masahiko Pakrasi, Himadri B. Ogawa, Teruo Aro, Eva-mari |
| Copyright Year | 2005 |
| Abstract | NDH (NADH-quinone oxidoreductase)-1 complexes in cyanobacteria have specific functions in respiration and cyclic electron flow as well as in active CO2 uptake. In order to isolate NDH-1 complexes and to study complex–complex interactions, several strains of Thermosynechococcus elongatus were constructed by adding a His-tag (histidine tag) to different subunits of NDH-1. Two strains with His-tag on CupA and NdhL were successfully used to isolate NDH-1 complexes by one-step Ni2+ column chromatography. BN (blue-native)/SDS/PAGE analysis of the proteins eluted from the Ni2+ column revealed the presence of three complexes with molecular masses of about 450, 300 and 190 kDa, which were identified by MS to be NDH-1L, NDH-1M and NDH-1S respectively, previously found in Synechocystis sp. PCC 6803. A larger complex of about 490 kDa was also isolated from the NdhL-His strain. This complex, designated ‘NDH-1MS’, was composed of NDH-1M and NDH-1S. NDH-1L complex was recovered from WT (wild-type) cells of T. elongatus by Ni2+ column chromatography. NdhF1 subunit present only in NDH-1L has a sequence of -HHDHHSHH- internally, which appears to have an affinity for the Ni2+ column. NDH-1S or NDH-1M was not recovered from WT cells by chromatography of this kind. The BN/SDS/PAGE analysis of membranes solubilized by a low concentration of detergent indicated the presence of abundant NDH-1MS, but not NDH-1M or NDH-1S. These results clearly demonstrated that NDH-1S is associated with NDH-1M in vivo. |
| Related Links | http://dx.doi.org/10.1042/bj20050390 |
| Ending Page | 520 |
| Page Count | 8 |
| Starting Page | 513 |
| File Format | |
| ISSN | 02646021 |
| e-ISSN | 14708728 |
| Journal | Biochemical Journal |
| Issue Number | Pt 2 |
| Volume Number | 390 |
| Language | English |
| Publisher | Portland Press Ltd. |
| Publisher Date | 2005-09-01 |
| Access Restriction | Open |
| Rights Holder | Portland Press Ltd. |
| Subject Keyword | Cell Biology Biochemistry Molecular Biology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Molecular Biology Biochemistry |
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