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Over‐expression and refolding of β‐subunit from the chloroplast ATP synthase
Author(s) -
Chen Zugen,
Wu Inmin,
L. Richter Mark,
Gegenheimer Peter
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80024-b
Subject(s) - atp synthase , chloroplast , protein subunit , chemistry , atp synthase gamma subunit , biochemistry , microbiology and biotechnology , biology , enzyme , atp hydrolysis , atpase , gene
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.