Open AccessExpression, purification and crystallization of the Atg5–Atg16 complex essential for autophagy
Author(s) -
Matsushita Minako,
Suzuki Nobuo N.,
Fujioka Yuko,
Ohsumi Yoshinori,
Inagaki Fuyuhiko
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106036232
Subject(s) - atg5 , atg12 , crystallography , autophagy , autophagosome , crystallization , crystal (programming language) , chemistry , stereochemistry , biochemistry , organic chemistry , apoptosis , computer science , programming language
Atg5 is a novel 34 kDa protein that is covalently modified by Atg12, a ubiquitin‐like modifier, and forms a complex with Atg16. The Atg12–Atg5–Atg16 complex localizes to autophagosome precursors and plays an essential role in autophagosome formation. Saccharomyces cerevisiae Atg5 in complex with the N‐terminal regions of Atg16 was expressed, purified and crystallized in four crystal forms. Forms I, II and III belong to space group P 2 1 , with unit‐cell parameters a = 66.3, b = 104.4, c = 112.1 Å, β = 92.1° (form I), a = 79.5, b = 101.4, c = 95.1 Å, β = 98.6° (form II) or a = 56.9, b = 101.2, c = 66.5 Å, β = 100.6° (form III). Form IV belongs to space group P 4 2 2 1 2, with unit‐cell parameters a = 73.3, c = 148.1 Å. Diffraction data were collected from all crystal forms and high‐resolution data to beyond 2.0 Å resolution were obtained from a form IV crystal.