Ca2+-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies

Helen Blanchard, Yunge Li, Miroslaw Cygler (Lead / Corresponding author), Cyril M. Kay, J. Simon C. Arthur, Peter L. Davies, John S. Elce

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

Abstract

The 21-kDa calcium-binding domain (VI) of the small subunit of rat calpain II has been expressed in Escherichia coli, purified, and crystallized. Two orthorhombic crystal forms have been obtained: space group P212121 with a = 50.3, b = 56.5, c = 141.3 Å; and space group C2221 with a = 69.4, b = 73.9, c = 157.4 Å. Diffraction data have been collected to 2.4 Å. Sedimentation equilibrium, dynamic light scattering, and gel-permeation chromatography indicate that domain VI exists as a homodimer in solution. In accordance with the protein's behavior in solution, each crystal form contains two molecules per asymmetric unit. Screening for heavy-atom derivatives is in progress. To decrease the sensitivity to mercurials and to aid in the search for useful derivatives, Cys-to-Ser mutants have been prepared, expressed, and crystallized.

Original languageEnglish
Pages (from-to)535-537
Number of pages3
JournalProtein Science
Volume5
Issue number3
DOIs
Publication statusPublished - Mar 1996

Keywords

  • Calcium-binding domain
  • Calpain
  • Crystallization
  • Homodimer
  • Smalt subunit

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