Crystallization and X-ray crystallographic analysis of m-calpain, a Ca2+-dependent protease.
The absolute requirement of Ca(2+) for proteolytic activity is a feature unique to the calpains, a family of heterodimeric cysteine proteases. Conditions are described which give rise to diffraction-quality crystals of m-calpain in two crystal forms, P1 and P2(1). Data have been collected from native crystals of m-calpain in both P1 and P2(1) forms, to 2.6 and 2.15 A, respectively. Selenomethionine-containing crystals have been grown in both forms, and anomalous data from the P2(1) selenomethionine enzyme provided the location of 17 of the 19 Se atoms in the protein.
The Protein Engineering Network of Centres of Excellence and Department of Biochemistry, Queen's University, Kingston, Ontario K7L 3N6, Canada.
C M
CM
Hosfield
Q
Q
Ye
J S
JS
Arthur
C
C
Hegadorn
D E
DE
Croall
J S
JS
Elce
Z
Z
Jia
D000818:Animals; D002154:Calpain / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D019281:Dimerization; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012645:Selenomethionine / Q000737:chemistry
D011994:Recombinant Proteins; D012645:Selenomethionine; D002154:Calpain