Crystallization of the coiled-coil domain of Atg16 essential for autophagy

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Nov 1;64(Pt 11):1046-8. doi: 10.1107/S1744309108031898. Epub 2008 Oct 31.

Abstract

Atg16 is a scaffold protein that interacts with Atg12-Atg5 protein conjugates via its N-terminal domain and self-assembles via its coiled-coil domain, thus forming a multimeric Atg12-Atg5-Atg16 complex that is essential for autophagy. The coiled-coil domain of Atg16 was expressed, purified and crystallized. The crystal belonged to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = 127.7, c = 77.8 A. Self-rotation functions and volume-to-weight ratio values suggested that the crystal may contain six molecules per asymmetric unit. Since the domain does not contain a methionine residue, selenomethionine-labelled crystals were prepared with a leucine-to-methionine substitution in the coiled-coil domain and these crystals were used for the collection of single-wavelength anomalous dispersion data to 2.5 A resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Autophagy*
  • Autophagy-Related Proteins
  • Carrier Proteins / chemistry*
  • Crystallization
  • Molecular Sequence Data
  • Multiprotein Complexes / chemistry
  • Protein Multimerization
  • Protein Structure, Tertiary*
  • Saccharomyces cerevisiae Proteins / chemistry*
  • X-Ray Diffraction

Substances

  • ATG16 protein, S cerevisiae
  • Autophagy-Related Proteins
  • Carrier Proteins
  • Multiprotein Complexes
  • Saccharomyces cerevisiae Proteins