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TIP47 protects mitochondrial membrane integrity and inhibits oxidative-stress-induced cell death
Authors:E. Hocsak  A. Szabo  E. Rapolti  Sz. Javor  F. Gallyas Jr.  A. Szigeti
Affiliation:a Department of Biochemistry and Medical Chemistry, University of Pécs, Pécs, Hungary
b Institute of Oncotherapy, University of Pécs, Pécs, Hungary
c Department of Surgical Research and Techniques, University of Pécs, Pécs, Hungary
Abstract:We found that overexpression of tail interacting protein of 47 kDa (TIP47), but not its truncated form (t-TIP47) protected NIH3T3 cells from hydrogen-peroxide-induced cell death, prevented the hydrogen-peroxide-induced mitochondrial depolarization determined by 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolylcarbocyanine iodide (JC1), while suppression of TIP47 in HeLa cells facilitated oxidative-stress-induced cell death. TIP47 was located to the cytoplasm of untreated cells, but some was associated to mitochondria in oxidative stress. Recombinant TIP47, but not t-TIP47 increased the mitochondrial membrane potential (Δψ), and partially prevented Ca2+ induced depolarization. It is assumed that TIP47 can bind to mitochondria in oxidative stress, and inhibit mitochondria mediated cell death by protecting mitochondrial membrane integrity.
Keywords:C-400, 5-(and-6)-carboxy-2&prime  ,7&prime  -dichlorodihydrofluorescein diacetate   DMEM, Dulbecco&rsquo  s modified Eagle&rsquo  s medium   dsiRNA, diced small interfering ribonucleic acid   ECL, electrochemiluminescence   FCS, fetal calf serum   FITC, fluorescein isothiocyanate   GST, glutathione S-transferase   IPTG, isopropyl-β-D-thiogalactopyranoside   JC-1, 5,5&prime  ,6,6&prime  -tetrachloro-1,1&prime  ,3,3&prime  -tetraethyl-benzimidazolylcarbocyanine iodide   Δψ, mitochondrial membrane potential   mPT, mitochondrial permeability transition   MTT, methylthiazolyldiphenyl-tetrazolium bromide   PBS, phosphate buffered saline   PCR, polymerase chain reaction   PI, propidium iodide   ROS, reactive oxygen species   TAG, triacylglycerol   TIP47, tail interacting protein of 47   kDa
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