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Role of the salt bridge between glutamate 546 and arginine 907 in preservation of autoinhibited form of Apaf-1.
Shakeri, Raheleh; Hosseinkhani, Saman; Los, Marek J; Davoodi, Jamshid; Jain, Mayur V; Cieslar-Pobuda, Artur; Rafat, Mehrdad; Ardestani, Sussan Kaboudanian.
Afiliación
  • Shakeri R; Institute of Biochemistry and Biophysics, Department of Biochemistry, University of Tehran, Tehran, Iran.
  • Hosseinkhani S; Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran. Electronic address: saman_h@modares.ac.ir.
  • Los MJ; Department of Clinical & Experimental Medicine (IKE), Division of Cell Biology, Integrative Regenerative Med. Center (IGEN), Linköping University, Linköping, Sweden; Department of Pathology, Pomeranian Medical University, Szczecin, Poland.
  • Davoodi J; Institute of Biochemistry and Biophysics, Department of Biochemistry, University of Tehran, Tehran, Iran.
  • Jain MV; Department of Clinical & Experimental Medicine (IKE), Division of Cell Biology, Integrative Regenerative Med. Center (IGEN), Linköping University, Linköping, Sweden.
  • Cieslar-Pobuda A; Department of Clinical & Experimental Medicine (IKE), Division of Cell Biology, Integrative Regenerative Med. Center (IGEN), Linköping University, Linköping, Sweden; Institute of Automatic Control, Silesian University of Technology, Gliwice, Poland.
  • Rafat M; Department of Biomedical Engineering, Linköping University, Linköping, Sweden.
  • Ardestani SK; Institute of Biochemistry and Biophysics, Department of Biochemistry, University of Tehran, Tehran, Iran. Electronic address: ardestany@ibb.ut.ac.ir.
Int J Biol Macromol ; 81: 370-4, 2015 Nov.
Article en En | MEDLINE | ID: mdl-26277751
Apaf-1, the key element of apoptotic mitochondrial pathway, normally exists in an auto-inhibited form inside the cytosol. WRD-domain of Apaf-1 has a critical role in the preservation of auto-inhibited form; however the underlying mechanism is unclear. It seems the salt bridges between WRD and NOD domains are involved in maintaining the inactive conformation of Apaf-1. At the present study, we have investigated the effect of E546-R907 salt bridge on the maintenance of auto-inhibited form of human Apaf-1. E546 is mutated to glutamine (Q) and arginine (R). Over-expression of wild type Apaf-1 and its E546Q and E546R variants in HEK293T cells does not induce apoptosis unlike - HL-60 cancer cell line. In vitro apoptosome formation assay showed that all variants are cytochrome c and dATP dependent to form apoptosome and activate endogenous procaspase-9 in Apaf-1-knockout MEF cell line. These results suggest that E546 is not a critical residue for preservation of auto-inhibited Apaf-1. Furthermore, the behavior of Apaf-1 variants for in vitro apoptosome formation in HEK293T cell is similar to exogenous wild type Apaf-1. Wild type and its variants can form apoptosome in HEK293T cell with different procaspase-3 processing pattern in the presence and absence of exogenous cytochrome c and dATP.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Arginina / Ácido Glutámico / Factor Apoptótico 1 Activador de Proteasas Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Int J Biol Macromol Año: 2015 Tipo del documento: Article País de afiliación: Irán Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Arginina / Ácido Glutámico / Factor Apoptótico 1 Activador de Proteasas Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Int J Biol Macromol Año: 2015 Tipo del documento: Article País de afiliación: Irán Pais de publicación: Países Bajos