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.
Hosfield
C M
CM
The Protein Engineering Network of Centres of Excellence and Department of Biochemistry, Queen's University, Kingston, Ontario K7L 3N6, Canada.
Ye
Q
Q
Arthur
J S
JS
Hegadorn
C
C
Croall
D E
DE
Elce
J S
JS
Jia
Z
Z
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