Crystallization and preliminary crystallographic study of a trypsin-resistant catalytic domain of human calcineurin.

ABSTRACT: Calcineurin, a Ca(2+)/calmodulin-dependent serine/threonine protein phosphatase, plays a key role in a number of cellular pathways, including T-cell activation, and is an important molecular target of the immunosuppressive drugs cyclosporin A and FK506. To understand the structural basis underlying the activation of calcineurin by calmodulin, X-ray crystallography was employed to solve the three-dimensional structure of the free calcineurin catalytic domain (residues 20-347 of the A subunit). To accomplish this, a bacterially expressed glutathione S-transferase (GST) fusion protein of the human calcineurin catalytic domain was first purified by GST-affinity chromatography. After limited digestion by trypsin, the catalytic domain (Cncat) was purified using anion-exchange and size-exclusion

SUBMITTER: Jin L 

PROVIDER: S-EPMC3374516 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature