Reassembly of Synechocystis sp. PCC 6803 F1‐ATPase from its over‐expressed subunits
- 3 April 1995
- journal article
- Published by Wiley in FEBS Letters
- Vol. 362 (2) , 171-174
- https://doi.org/10.1016/0014-5793(95)00238-5
Abstract
Subunits α, β, and γ of the F1‐part of cyanobacterial F0F1‐ATPase have been cloned into expression vectors. Overexpressed subunit β was found soluble in the cytoplasmic fraction of Escherichia coli cells under appropriate culture and induction conditions and was purified from cell extracts. Recombinant α and γ subunits precipitated into inclusion bodies and had to be solubilized, purified and refolded. The correct folding and functional integrity of the α and β subunits was monitored by their ability to bind nucleotides. Active cyanobacterial F1‐ATPase was assembled from its purified subunits α, β, γ, δ and ϵ. The reassembled enzyme reconstituted ATP synthesis in F1‐depleted thylakoid membranes of Synechocystis sp. PCC 6803 and hydrolyzed ATP.Keywords
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