title: Calpain cleaves methionine aminopeptidase-2 in a rat model of ischemia/reperfusion.
abstract: Ischemic stroke results in multiple injurious signals within a cell including dysregulation of calcium homeostasis. Consequently, there is an increase in the enzymatic activity of the calpains, calcium dependent proteases that are thought to contribute to neuronal injury. In addition, cellular stress due to ischemia/reperfusion also triggers a decrease in protein translation through activation of the unfolded protein response (UPR). In the present study we found that methionine aminopeptidase 2 (MetAP2), a critical component of the translation initiation complex, is a calpain substrate. In vitro calpain assays demonstrated that while MetAP2 has autoproteolytic activity, calpain also produces a stable proteolytic fragment at 50kDa using recombinant MetAP2. This 50kDa fragment, in addition to a 57kDa fragment was present in in vitro digestions of rat brain homogenates. Production of these fragments was inhibited by calpastatin, the endogenous and specific inhibitor of calpain. Using an in vivo middle cerebral artery occlusion (MCAO) model only the 57kDa fragment of MetAP2 was observed. These data suggest that calpain activation in stroke may regulate MetAP2-mediated protein translation giving calpains a larger role in the cellular stress response than previously determined.
authors.affiliation: Department of Gerontology, University of Kentucky, Lexington, KY, USA.
authors.forename: Tiffanie
authors.initials: T
authors.lastname: Clinkinbeard
authors.forename: Sarbani
authors.initials: S
authors.lastname: Ghoshal
authors.forename: Susan
authors.initials: S
authors.lastname: Craddock
authors.forename: L
authors.initials: L
authors.lastname: Creed Pettigrew
authors.forename: Rodney P
authors.initials: RP
authors.lastname: Guttmann
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D002154:Calpain / Q000378:metabolism*; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D051381:Rats; D015427:Reperfusion Injury / Q000378:metabolism*
chemical_list: D000626:Aminopeptidases; C106541:methionine aminopeptidase 2; D002154:Calpain; D008666:Metalloendopeptidases