MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. EMBO Rep. (1469-221X) 2014 imex curation Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu Accepted 2014-MAY-26 by MARTA EMBO Rep. (1469-221X) 2014 Only protein-protein interactions imex curation Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. EMBO Rep. (1469-221X) 2014 imex curation Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu Accepted 2014-MAY-26 by MARTA EMBO Rep. (1469-221X) 2014 Only protein-protein interactions imex curation Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. EMBO Rep. (1469-221X) 2014 imex curation Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu Accepted 2014-MAY-26 by MARTA EMBO Rep. (1469-221X) 2014 Only protein-protein interactions imex curation Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy. EMBO Rep. (1469-221X) 2014 imex curation Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro cosedimentation cosedimentation western blot western blot Immuno blot Immuno blot Fedorowicz M.A., de Vries-Schneider R.L., Rüb C., Becker D., Huang Y., Zhou C., Alessi Wolken D.M., Voos W., Liu Y., Przedborski S. sp30@columbia.edu Accepted 2014-MAY-26 by MARTA EMBO Rep. (1469-221X) 2014 Only protein-protein interactions imex curation tim23_human Mitochondrial import inner membrane translocase subunit Tim23 TIMM23 TIM23 protein protein human Homo sapiens Human MEGGGGSGNKTTGGLAGFFGAGGAGYSHADLAGVPLTGMNPLSPYLNVDPRYLVQDTDEFILPTGANKTRGRFELAFFTIGGCCMTGAAFGAMNGLRLGLKETQNMAWSKPRNVQILNMVTRQGALWANTLGSLALLYSAFGVIIEKTRGAEDDLNTVAAGTMTGMLYKCTGGLRGIARGGLTGLTLTSLYALYNNWEHMKGSLLQQSL C9A1BBEC703B6838 tom20_human Mitochondrial import receptor subunit TOM20 homolog Outer mitochondrial membrane receptor Tom20 TOMM20 KIAA0016 Mitochondrial 20 kDa outer membrane protein protein protein human Homo sapiens Human MVGRNSAIAAGVCGALFIGYCIYFDRKRRSDPNFKNRLRERRKKQKLAKERAGLSKLPDLKDAEAVQKFFLEEIQLGEELLAQGEYEKGVDHLTNAIAVCGQPQQLLQVLQQTLPPPVFQMLLTKLPTISQRIVSAQSLAEDDVE DIP protein Q15388 original sequence version: 139 5153BD25AC5D9B3A prkn_human E3 ubiquitin-protein ligase parkin PRKN PARK2 Parkin RBR E3 ubiquitin-protein ligase Parkinson juvenile disease protein 2 protein protein human Homo sapiens Human MIVFVRFNSSHGFPVEVDSDTSIFQLKEVVAKRQGVPADQLRVIFAGKELRNDWTVQNCDLDQQSIVHIVQRPWRKGQEMNATGGDDPRNAAGGCEREPQSLTRVDLSSSVLPGDSVGLAVILHTDSRKDSPPAGSPAGRSIYNSFYVYCKGPCQRVQPGKLRVQCSTCRQATLTLTQGPSCWDDVLIPNRMSGECQSPHCPGTSAEFFFKCGAHPTSDKETSVALHLIATNSRNITCITCTDVRSPVLVFQCNSRHVICLDCFHLYCVTRLNDRQFVHDPQLGYSLPCVAGCPNSLIKELHHFRILGEEQYNRYQQYGAEECVLQMGGVLCPRPGCGAGLLPEPDQRKVTCEGGNGLGCGFAFCRECKEAYHEGECSAVFEASGTTTQAYRVDERAAEQARWEAASKETIKKTTKPCPRCHVPVEKNGGCMHMKCPQPQCRLEWCWNCGCEWNRVCMGDHWFDV DIP protein O60260 original sequence version: 99 DIP protein O60260-1 original sequence version: 145 DIP protein O60260-1 original sequence version: 158 DIP protein O60260 original sequence version: 157 9A8BB802A3FC84C3 pink1_human Serine/threonine-protein kinase PINK1, mitochondrial PINK1 BRPK PTEN-induced putative kinase protein 1 protein protein human Homo sapiens Human MAVRQALGRGLQLGRALLLRFTGKPGRAYGLGRPGPAAGCVRGERPGWAAGPGAEPRRVGLGLPNRLRFFRQSVAGLAARLQRQFVVRAWGCAGPCGRAVFLAFGLGLGLIEEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEYLIGQSIGKGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFPLAIKMMWNISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNIIRVLRAFTSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNTPSPRLAAMMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADESIGLQLPFSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVNPFYGQGKAHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASKRPSARVAANVLHLSLWGEHILALKNLKLDKMVGWLLQQSAATLLANRLTEKCCVETKMKMLFLANLECETLCQAALLLCSWRAAL 721FE01F63263A64 pink1-timm23 4 8 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level colocalization colocalization f1a Here, we transiently transfected HeLa cells with HA‐tagged human PINK1 and treated them with either MG132, valinomycin or both for 8 h. Incubation with MG132 was associated with a marked increase in PINK152, not only in the crude mitochondrial fraction but also in the cytosolic fraction (Fig 1A), confirming that PINK152 is present in both cellular compartments [12], [13], [14], [15], [16]. Conversely, incubation with valinomycin was associated with a decrease in PINK152 in both subcellular fractions while there was an increase in PINK163 in the mitochondrial fractions (Fig 1A). Finally, the combination of valinomycin and MG132 decreased the content of PINK152 in both subcellular fractions compared to MG132 alone (Fig 1A). pink1-tomm20-1 2 8 immunostaining immunostaining immunostaining immunostaining unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell 6 immunostaining immunostaining immunostaining immunostaining unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f1b As illustrated by immunocytochemistry (Fig 1B), under basal conditions (DMSO control), there was marginal co‐localization between PINK1 and the mitochondrial marker TOM20 (co‐localization coefficient c = 0.16). In contrast, after 2 h exposure to valinomycin, there was a greater co‐localization between PINK1 and TOM20 (c = 0.58), with PINK1 immunoreactivity displaying a prominent punctate distribution. Following the incubation of cells with MG132, we saw cytosolic PINK1+ aggregates, many of which were TOM20−. Consequently, the mitochondrial co‐localization was limited (c = 0.32), again suggesting that a significant portion of PINK152 is localized in the cytosol. park2-pink1-1 3 8 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2a pink1-park2 3 7 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association f2b pink1-park2-1 1 7 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association f2b