Subunit topology of the Rhodococcus proteasome

FEBS Letters

Dolenc, Iztok²  Baumeister, Wolfgang²  De Mot, René¹  Tamura, Tomohiro²  Zühl, Frank²  Cejka, Zdenka²  Nagy, István¹ 

[1] F.A. Janssens Laboratory of Genetics, Catholic University of Leuven, B-3001 Heverlee, Belgium;Max-Planck-Institute for Biochemistry, D-82152 Martinsried, Germany

关键词: Proteasome;    Proteolysis;    Rhodococcus;    Gene expression;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    DMSO;    dimethylsulfoxide;    DTT;    dithiothreitol;    AMC;    7-amino-4-methylcoumarin;    Suc;    succinyl;    Z;    benzoxycarbonyl;    Boc;    N-t-butoxycarbonyl;    Bz;    benzonyl;    MW;    molecular weight;   

DOI  :  10.1016/S0014-5793(96)01403-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The 20S proteasome, isolated from the nocardioform actinomycete Rhodococcus erythropolis strain NI86/21, is built from two α-type and two β-type subunits. In order to probe the subunit topology, we have set up an expression system which allows coexpression of the genes encoding the α- and β-subunits in all possible combinations. The four respective constructs obtained yielded fully assembled and proteolytically active proteasomes. Biochemical, kinetic and electron microscopy analysis allow us to rule out several of the models which were originally envisaged for the subunit topology of the Rhodococcus proteasome. The experiments further indicate that the assembly pathways of the Rhodococcus and of the Thermoplasma proteasome differ in some important details.

【 授权许可】

Unknown