Frq2 from Drosophila melanogaster: cloning, expression, purification, crystallization and preliminary X-ray analysis

Acta Crystallogr F Struct Biol Commun. 2014 Apr;70(Pt 4):530-4. doi: 10.1107/S2053230X14005408. Epub 2014 Mar 25.

Abstract

Drosophila melanogaster contains two calcium-binding proteins, Frq1 and Frq2, in the nervous system that control the number of synapses and the probability of release. To understand the differential function of the two proteins, whose sequence is only 5% dissimilar, the crystal structures of Frq1 and Frq2 are needed. Here, the cloning, expression, purification, crystallization and preliminary crystallographic analysis of Frq2 are presented. The full-length protein was purified using a two-step chromatographic procedure. Two different diffracting crystal forms were obtained using a progressive streak-seeding method and detergents.

Keywords: Drosophila melanogaster; Frq2; calcium-binding proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium-Binding Proteins / chemistry*
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / isolation & purification*
  • Cloning, Molecular*
  • Crystallization / methods*
  • Crystallography, X-Ray / methods*
  • Drosophila Proteins / chemistry*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / isolation & purification*
  • Drosophila melanogaster / metabolism*

Substances

  • Calcium-Binding Proteins
  • Drosophila Proteins
  • FRQ2 protein, Drosophila