Expression of mammalian mitochondrial F1-ATPase in Escherichia coli depends on two chaperone factors, AF1 and AF2

Toshiharu Suzuki; Naoya Iida; Junko Suzuki; Yasunori Watanabe; Toshiya Endo; Toru Hisabori; Masasuke Yoshida

doi:10.1002/2211-5463.12143

Expression of mammalian mitochondrial F₁-ATPase in Escherichia coli depends on two chaperone factors, AF1 and AF2

FEBS Open Bio. 2016 Oct 25;6(12):1267-1272. doi: 10.1002/2211-5463.12143. eCollection 2016 Dec.

Authors

Toshiharu Suzuki¹, Naoya Iida², Junko Suzuki³, Yasunori Watanabe³, Toshiya Endo³, Toru Hisabori⁴, Masasuke Yoshida³

Affiliations

¹ Faculty of Science and Engineering Waseda University Tokyo Japan; Department of Molecular Bioscience Kyoto-Sangyo University Kyoto Japan; Chemical Resources Laboratory Tokyo Institute of Technology Yokohama Japan; Present address: Department of Applied Chemistry School of Engineering The University of Tokyo Hongo, Bunkyo-ku Tokyo 113-8656 Japan.
² Faculty of Science and Engineering Waseda University Tokyo Japan.
³ Department of Molecular Bioscience Kyoto-Sangyo University Kyoto Japan.
⁴ Chemical Resources Laboratory Tokyo Institute of Technology Yokohama Japan.

Abstract

F₁-ATPase (F₁) is a multisubunit water-soluble domain of F_oF_1- ATP synthase and is a rotary enzyme by itself. Earlier genetic studies using yeast suggested that two factors, Atp11p and Atp12p, contribute to F₁ assembly. Here, we show that their mammalian counterparts, AF1 and AF2, are essential and sufficient for efficient production of recombinant bovine mitochondrial F₁ in Escherichia coli cells. Intactness of the function and conformation of the E. coli-expressed bovine F₁ was verified by rotation analysis and crystallization. This expression system opens a way for the previously unattempted mutation study of mammalian mitochondrial F₁.

Keywords: F1‐ATPase; FoF1‐ATP synthase; molecular chaperone.