Affinity purification of human m-calpain through an intrinsically disordered inhibitor, calpastatin
  • Metaadatok
Tartalom: http://real.mtak.hu/75152/
Archívum: MTA Könyvtár
Gyűjtemény: Status = Published


Type = Article
Cím:
Affinity purification of human m-calpain through an intrinsically disordered inhibitor, calpastatin
Létrehozó:
Nguyen, H. H.
Váradi, Mihály
Tompa, PĂ©ter
Pauwels, K.
Kiadó:
Public Library of Science (PLoS)
Dátum:
2017
Téma:
QH3011 Biochemistry / biokémia
QH3020 Biophysics / biofizika
Tartalmi leírás:
Calpains are calcium-activated proteases that have biomedical and biotechnological potential. Their activity is tightly regulated by their endogenous inhibitor, calpastatin that binds to the enzyme only in the presence of calcium. Conventional approaches to purify calpain comprise multiple chromatographic steps, and are labor-intensive, leading to low yields. Here we report a new purification procedure for the human m-calpain based on its reversible calcium-mediated interaction with the intrinsically disordered calpastatin. We exploit the specific binding properties of human calpastatin domain 1 (hCSD1) to physically capture human m-calpain from a complex biological mixture. The dissociation of the complex is mediated by chelating calcium, upon which heterodimeric calpain elutes while hCSD1 remains immobilized onto the stationary phase. This novel affinity-based purification was compared to the conventional multistep purification strategy and we find that it is robust, it yields a homogeneous preparation, it can be scaled up easily and it rests on a non-disruptive step that maintains close to physiological conditions that allow further biophysical and functional studies. © 2017 Nguyen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Nyelv:
angol
Típus:
Article
PeerReviewed
info:eu-repo/semantics/article
Formátum:
text
Azonosító:
Nguyen, H. H. and Váradi, Mihály and Tompa, Péter and Pauwels, K. (2017) Affinity purification of human m-calpain through an intrinsically disordered inhibitor, calpastatin. PLOS ONE, 12 (3). pp. 1-15. ISSN 1932-6203
Kapcsolat:
https://doi.org/10.1371/journal.pone.0174125
MTMT:3211114; doi:10.1371/journal.pone.0174125