Expression of an archaeal chaperonin in E. coli: formation of homo‐ (α, β) and hetero‐oligomeric (α + β) thermosome complexes

FEBS Letters

Baumeister, Wolfgang¹  Klumpp, Martin¹  Waldmann, Thomas¹  Nitsch, Michael¹ 

[1] Max Planck Institut für Biochemie, Am Klopferspitz 18a, 82152 Martinsried, Germany

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;   

DOI  :  10.1016/0014-5793(95)01248-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown