Expression of an archaeal chaperonin in E. coli: formation of homo‐ (α, β) and hetero‐oligomeric (α + β) thermosome complexes
[摘要] Co-expression of the two genes encoding the α- and β-subunit of the Thermoplasma acidophilum thermosome in Escherichia coli yielded fully assembled hetero-oligomeric complexes (α + β). Surprisingly, also separate expression of both genes resulted in formation of hexadecameric complexes (α, β) in the bacterial cytoplasm. On electron micrographs these complexes were indistinguishable from each other and from the native thermosome. The recombinant a-complex as well as the native thermosome could be reconstituted in vitro from their dissociated subunits in the presence of Mg-ATP.
[发布日期] [发布机构]
[效力级别] [学科分类] 生物化学/生物物理
[关键词] Thermosome;Molecular chaperone;Gene expression;Archaebacteria;Thermoplasma acidophilum;Electron microscopy;AMPPNP;5′-adenylimido-diphosphate;ATPγS;adenosine 5′-O-(3-thiotriphosphate);CCT;chaperonin containing TCP-1;IPTG;isopropyl-β-d-thiogalactopyranoside;OD600;optical density measured at 600 nm;PCR;polymerase chain reaction;pl;isoelectric point;PMSF;phenylmethanesulfonyl fluoride;TF55/56;thermophilic factor of 55/56 kDa;TRiC;TCP-1 ring complex [时效性]
