Expression, purification and crystallization of the Atg5-Atg16 complex essential for autophagy

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Oct 1;62(Pt 10):1021-3. doi: 10.1107/S1744309106036232. Epub 2006 Sep 30.

Abstract

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 P2(1), with unit-cell parameters a = 66.3, b = 104.4, c = 112.1 A, beta = 92.1 degrees (form I), a = 79.5, b = 101.4, c = 95.1 A, beta = 98.6 degrees (form II) or a = 56.9, b = 101.2, c = 66.5 A, beta = 100.6 degrees (form III). Form IV belongs to space group P4(2)2(1)2, with unit-cell parameters a = 73.3, c = 148.1 A. Diffraction data were collected from all crystal forms and high-resolution data to beyond 2.0 A resolution were obtained from a form IV crystal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Carrier Proteins / chemistry*
  • Carrier Proteins / isolation & purification
  • Carrier Proteins / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / metabolism
  • Ubiquitin-Protein Ligases

Substances

  • ATG16 protein, S cerevisiae
  • ATG5 protein, S cerevisiae
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Carrier Proteins
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin-Protein Ligases