TY - JOUR
T1 - Ca2+-Binding domain VI of rat calpain is a homodimer in solution
T2 - Hydrodynamic, crystallization and preliminary X-ray diffraction studies
AU - Blanchard, Helen
AU - Li, Yunge
AU - Cygler, Miroslaw
AU - Kay, Cyril M.
AU - Arthur, J. Simon C.
AU - Davies, Peter L.
AU - Elce, John S.
PY - 1996/3
Y1 - 1996/3
N2 - 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.
AB - 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.
KW - Calcium-binding domain
KW - Calpain
KW - Crystallization
KW - Homodimer
KW - Smalt subunit
UR - http://www.scopus.com/inward/record.url?scp=0029670455&partnerID=8YFLogxK
U2 - 10.1002/pro.5560050317
DO - 10.1002/pro.5560050317
M3 - Article
C2 - 8868491
AN - SCOPUS:0029670455
SN - 0961-8368
VL - 5
SP - 535
EP - 537
JO - Protein Science
JF - Protein Science
IS - 3
ER -