Expression in Escherichia coli of a sub‐gene encoding the lipoyl domain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus

FEBS Letters

Packman, Leonard C.¹  Perham, Richard N.¹  Dardel, Frédéric¹ 

[1] Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 IQW, UK

关键词: Lipoyl domain;    Pyruvate dehydrogenase complex;    Lipoylation;    NMR spectroscopy;    DTT;    dithiothreitol;    PDH;    pyruvate dehydrogenase;    PTH;    phenyl thiohydantoin;    SDS;    sodium dodecylsulphate;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/0014-5793(90)80249-I

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A sub-gene encoding the lipoyl domain (residues 1–85) of the lipoate acetyltransferase chain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus was over-expressed in Escherichia coli. Approx. 80% of the domain was unlipoylated but most of the remainder was correctly lipoylated on Lys-42 and could be reductively acetylated by the B stearothermophilus enzyme complex. A small proportion (approx. 4%) of the domain carried an aberrant substituent, possibly an octanoyl group, on Lys-42. The 400 MHz ¹H NMR spectra of the lipoylated and unlipoylated domains were essentially identical and closely resembled that of the native lipoyl domain.

【 授权许可】

Unknown