Concerted Multi-Pronged Attack by Calpastatin to Occlude the Catalytic Cleft of Heterodimeric Calpains
The Ca{sup 2+}-dependent cysteine proteases, calpains, regulate cell migration, cell death, insulin secretion, synaptic function and muscle homeostasis. Their endogenous inhibitor, calpastatin, consists of four inhibitory repeats, each of which neutralizes an activated calpain with exquisite specificity and potency. Despite the physiological importance of this interaction, the structural basis of calpain inhibition by calpastatin is unknown. Here we report the 3.0{angstrom} structure of Ca{sup 2+}-bound m-calpain in complex with the first calpastatin repeat, both from rat, revealing the mechanism of exclusive specificity. The structure highlights the complexity of calpain activation by Ca{sup 2+}, illustrating key residues in a peripheral domain that serve to stabilize the protease core on Ca{sup 2+} binding. Fully activated calpain binds ten Ca{sup 2+} atoms, resulting in several conformational changes allowing recognition by calpastatin. Calpain inhibition is mediated by the intimate contact with three critical regions of calpastatin. Two regions target the penta-EF-hand domains of calpain and the third occupies the substrate-binding cleft, projecting a loop around the active site thiol to evade proteolysis.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
- Sponsoring Organization:
- Doe - Office Of Science
- DOE Contract Number:
- DE-AC02-98CH10886
- OSTI ID:
- 980077
- Report Number(s):
- BNL-92995-2010-JA; TRN: US201015%%1462
- Journal Information:
- Nature, Vol. 456
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
ATOMS
CONFORMATIONAL CHANGES
CYSTEINE
DEATH
FUNCTIONS
HOMEOSTASIS
INHIBITION
INSULIN
MIGRATION
MUSCLES
PROTEOLYSIS
RESIDUES
SECRETION
SPECIFICITY
TARGETS
THIOLS
national synchrotron light source