Live cell interactome of the human voltage dependent anion channel 3 (VDAC3) revealed in HeLa cells by affinity purification tag technique

摘要: In higher eukaryotes three different VDAC genes encode homologous proteins which do not show the same activity. VDAC1 and VDAC2 isoforms have been characterized while VDAC3 isoform is still elusive. To explore protein interactions, we established a stable cell line expressing fluorescent dual-tagged construct. This clone expresses amount of VDAC3. Live imaging shows that localizes in mitochondria. Proteins interacting with separated by tandem-affinity purification 2-D gel electrophoresis identified mass spectrometry. list putative proteins, there are cytosolic, mitochondrial, cytoskeletal ER proteins. Coherent pathways like redox homeostasis, response to stress, formation/rearrangement disulfide bonds, unfolded or folding found be related clusters this experiment. The associated has validated immunoprecipitation experiments utilizing specific antibodies. Likely biological pathological processes analyzed. Cytosolic include tubulins stress sensors, chaperones proteasome components, redox-mediating enzymes such as disulphide isomerase. overall picture points role for mediator organization complexes regulator traffic misfolded non-folded evoked from stimuli.