Open AccessTwo‐dimensional crystallization of intact F‐ATP synthase isolated from bovine heart mitochondria
Author(s) -
Maeda Shintaro,
ShinzawaItoh Kyoko,
Mieda Kaoru,
Yamamoto Mami,
Nakashima Yumiko,
Ogasawara Yumi,
Jiko Chimari,
Tani Kazutoshi,
Miyazawa Atsuo,
Gerle Christoph,
Yoshikawa Shinya
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113029072
Subject(s) - atp synthase , crystallization , mitochondrion , chemistry , biochemistry , enzyme , organic chemistry
Mitochondrial F‐ATP synthase produces the majority of ATP for cellular functions requiring free energy. The structural basis for proton motive force‐driven rotational catalysis of ATP formation in the holoenzyme remains to be determined. Here, the purification and two‐dimensional crystallization of bovine heart mitochondrial F‐ATP synthase are reported. Two‐dimensional crystals of up to 1 µm in size were grown by dialysis‐mediated detergent removal from a mixture of decylmaltoside‐solubilized 1,2‐dimyristoyl‐ sn ‐glycero‐3‐phosphocholine and F‐ATP synthase against a detergent‐free buffer. A projection map calculated from an electron micrograph of a negatively stained two‐dimensional crystal revealed unit‐cell parameters of a = 185.0, b = 170.3 Å, γ = 92.5°.