Over‐expression and refolding of β‐subunit from the chloroplast ATP synthase

FEBS Letters

L. Richter, Mark¹  Chen, Zugen¹  Wu, Inmin¹  Gegenheimer, Peter¹ 

[1] Department of Biochemistry, Botany and Molecular Genetics Program, The University of Kansas, Lawrence, Kansas, USA

关键词: ATP synthase;    Over-expressed (chloroplast) β subunit;    CF1;    chloroplast couling factor 1;    TNP-ATP;    2′;    3′-O(2;    4;    6-trinitrophenyl)ATP;    Tricine;    N-(2-hydroxy-1;    1-bis(hydroxymethyl)ethyl) glycine;    Tris;    tris(hydroxymethyl)aminomethane;    IPTG;    isopropyl-β-D-thiogalactoside;   

DOI  :  10.1016/0014-5793(92)80024-B

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We established a bacterial system for high-level over-expression of the spinach chloroplast atpB gene which encodes the ATP Synthase β subunit. Upon induction, atpB was expressed as at least 50% to 70% of total cell protein. Although the over-expressed β polypeptide formed insoluble inclusion bodies, more than fifty percent of it was restored to a functional form by solubilizing the inclusion bodies with 4 M urea and slowly removing the urea by stepwise dialysis. The resulting β subunit exhibited specific and selective nucleotide binding properties identical to those of the native β subunit.

【 授权许可】

Unknown