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

Journal Article


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.

Publication Date


  • 1996

Citation


  • Blanchard, H., Li, Y., Cygler, M., Kay, C. M., Arthur, J. S. C., Davies, P. L., & Elce, J. S. (1996). Ca2+-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies. Protein Science, 5(3), 535-537. doi:10.1002/pro.5560050317

Scopus Eid


  • 2-s2.0-0029670455

Start Page


  • 535

End Page


  • 537

Volume


  • 5

Issue


  • 3

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.

Publication Date


  • 1996

Citation


  • Blanchard, H., Li, Y., Cygler, M., Kay, C. M., Arthur, J. S. C., Davies, P. L., & Elce, J. S. (1996). Ca2+-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies. Protein Science, 5(3), 535-537. doi:10.1002/pro.5560050317

Scopus Eid


  • 2-s2.0-0029670455

Start Page


  • 535

End Page


  • 537

Volume


  • 5

Issue


  • 3