Prediction of the binding site on E1 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus

FEBS Letters

Perham, Richard N¹  Jung, Hyo-Il¹ 

[1] Cambridge Centre for Molecular Recognition, Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK

关键词: Pyruvate dehydrogenase;    Homology modelling;    Multienzyme complex;    Alanine-scanning mutagenesis;    Protein–protein interaction;    PDH;    pyruvate dehydrogenase;    BCDH;    branched-chain 2-oxo acid dehydrogenase;    E1;    pyruvate decarboxylase (EC 1.2.4.1);    E2;    dihydrolipoyl acetyltransferase (EC 2.3.1.12);    E3;    dihydrolipoyl dehydrogenase (EC 1.8.1.4);    PSBD;    peripheral subunit-binding domain;    LD;    lipoyl domain;    CD;    catalytic domain;    ThDD;    thrombin-cleavable di-domain;    SPR;    surface plasmon resonance;   

DOI  :  10.1016/S0014-5793(03)01245-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The β-subunit (E1β) of the pyruvate decarboxylase (E1, α₂β₂) component of the Bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of Pseudomonas putida and Homo sapiens. Based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of Glu285 and the hydrophobic side chain of Phe324 are of particular importance in the interaction with the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the complex. These results help to identify the site of interaction on the E1β subunit and are consistent with thermodynamic evidence of a mixture of electrostatic and hydrophobic interactions being involved.

【 授权许可】

Unknown