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 Muscle Lim Protein isoform negatively regulates striated muscle actin dynamics and differentiation. Muscle Lim Protein isoform negatively regulates striated muscle actin dynamics and differentiation. FEBS J. (1742-464X) 2014 imex curation Vafiadaki E., Arvanitis D.A., Papalouka V., Terzis G., Roumeliotis T.I., Spengos K., Garbis S.D., Manta P., Kranias E.G., Sanoudou D. dsanoudo@enders.tch.harvard.edu 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western dsanoudo@enders.tch.harvard.edu Accepted 2014-MAY-26 by LUANA 2014 imex curation Muscle Lim Protein isoform negatively regulates striated muscle actin dynamics and differentiation. Muscle Lim Protein isoform negatively regulates striated muscle actin dynamics and differentiation. FEBS J. (1742-464X) 2014 imex curation Vafiadaki E., Arvanitis D.A., Papalouka V., Terzis G., Roumeliotis T.I., Spengos K., Garbis S.D., Manta P., Kranias E.G., Sanoudou D. dsanoudo@enders.tch.harvard.edu 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot dsanoudo@enders.tch.harvard.edu Accepted 2014-MAY-26 by LUANA 2014 imex curation csrp3_human Cysteine and glycine-rich protein 3 CLP Cardiac LIM protein Cysteine-rich protein 3 LIM domain protein, cardiac MLP Muscle LIM protein CSRP3 protein protein human Homo sapiens Human MPNWGGGAKCGACEKTVYHAEEIQCNGRSFHKTCFHCMACRKALDSTTVAAHESEIYCKVCYGRRYGPKGIGYGQGAGCLSTDTGEHLGLQFQQSPKPARSVTTSNPSKFTAKFGESEKCPRCGKSVYAAEKVMGGGKPWHKTCFRCAICGKSLESTNVTDKDGELYCKVCYAKNFGPTGIGFGGLTQQVEKKE FDB6E4F8D258C35F actn2_human Alpha-actinin-2 ACTN2 Alpha-actinin skeletal muscle isoform 2 F-actin cross-linking protein protein protein human Homo sapiens Human MNQIEPGVQYNYVYDEDEYMIQEEEWDRDLLLDPAWEKQQRKTFTAWCNSHLRKAGTQIENIEEDFRNGLKLMLLLEVISGERLPKPDRGKMRFHKIANVNKALDYIASKGVKLVSIGAEEIVDGNVKMTLGMIWTIILRFAIQDISVEETSAKEGLLLWCQRKTAPYRNVNIQNFHTSWKDGLGLCALIHRHRPDLIDYSKLNKDDPIGNINLAMEIAEKHLDIPKMLDAEDIVNTPKPDERAIMTYVSCFYHAFAGAEQAETAANRICKVLAVNQENERLMEEYERLASELLEWIRRTIPWLENRTPEKTMQAMQKKLEDFRDYRRKHKPPKVQEKCQLEINFNTLQTKLRISNRPAFMPSEGKMVSDIAGAWQRLEQAEKGYEEWLLNEIRRLERLEHLAEKFRQKASTHETWAYGKEQILLQKDYESASLTEVRALLRKHEAFESDLAAHQDRVEQIAAIAQELNELDYHDAVNVNDRCQKICDQWDRLGTLTQKRREALERMEKLLETIDQLHLEFAKRAAPFNNWMEGAMEDLQDMFIVHSIEEIQSLITAHEQFKATLPEADGERQSIMAIQNEVEKVIQSYNIRISSSNPYSTVTMDELRTKWDKVKQLVPIRDQSLQEELARQHANERLRRQFAAQANAIGPWIQNKMEEIARSSIQITGALEDQMNQLKQYEHNIINYKNNIDKLEGDHQLIQEALVFDNKHTNYTMEHIRVGWELLLTTIARTINEVETQILTRDAKGITQEQMNEFRASFNHFDRRKNGLMDHEDFRACLISMGYDLGEAEFARIMTLVDPNGQGTVTFQSFIDFMTRETADTDTAEQVIASFRILASDKPYILAEELRRELPPDQAQYCIKRMPAYSGPGSVPGALDYAAFSSALYGESDL DIP protein P35609 original sequence version: 118 DIP protein P35609 original sequence version: 117 7F612C1C3B3E2299 myod1_human Myoblast determination protein 1 Myogenic factor 3 MYOD1 MYF3 MYOD BHLHC1 Class C basic helix-loop-helix protein 1 protein protein human Homo sapiens Human MELLSPPLRDVDLTAPDGSLCSFATTDDFYDDPCFDSPDLRFFEDLDPRLMHVGALLKPEEHSHFPAAVHPAPGAREDEHVRAPSGHHQAGRCLLWACKACKRKTTNADRRKAATMRERRRLSKVNEAFETLKRCTSSNPNQRLPKVEILRNAIRYIEGLQALLRDQDAAPPGAAAAFYAPGPLPPGRGGEHYSGDSDASSPRSNCSDGMMDYSGPPSGARRRNCYEGAYYNEAPSEPRPGKSAAVSSLDCLSSIVERISTESPAAPALLLADVPSESPPRRQEAAAPSEGESSGDPTQSPDAAPQCPAGANPNPIYQVL 75E624D2ED5B0B33 cof2_human Cofilin-2 CFL2 Cofilin, muscle isoform protein protein human Homo sapiens Human MASGVTVNDEVIKVFNDMKVRKSSTQEEIKKRKKAVLFCLSDDKRQIIVEEAKQILVGDIGDTVEDPYTSFVKLLPLNDCRYALYDATYETKESKKEDLVFIFWAPESAPLKSKMIYASSKDAIKKKFTGIKHEWQVNGLDDIKDRSTLGEKLGGNVVVSLEGKPL DIP protein Q9Y281 original sequence version: 121 48B6CDCCAE9FE1CC telt_human Telethonin Titin cap protein TCAP protein protein human Homo sapiens Human MATSELSCEVSEENCERREAFWAEWKDLTLSTRPEEGCSLHEEDTQRHETYHQQGQCQVLVQRSPWLMMRMGILGRGLQEYQLPYQRVLPLPIFTPAKMGATKEEREDTPIQLQELLALETALGGQCVDRQEVAEITKQLPPVVPVSKPGALRRSLSRSMSQEAQRG DIP protein O15273 original sequence version: null DIP protein O15273 original sequence version: 117 DIP protein O15273 original sequence version: 89 A3B0E27D8C84F6C5 myog_human Myogenin MYOG BHLHC3 MYF4 Class C basic helix-loop-helix protein 3 Myogenic factor 4 protein protein human Homo sapiens Human MELYETSPYFYQEPRFYDGENYLPVHLQGFEPPGYERTELTLSPEAPGPLEDKGLGTPEHCPGQCLPWACKVCKRKSVSVDRRRAATLREKRRLKKVNEAFEALKRSTLLNPNQRLPKVEILRSAIQYIERLQALLSSLNQEERDLRYRGGGGPQPGVPSECSSHSASCSPEWGSALEFSANPGDHLLTADPTDAHNLHSLTSIVDSITVEDVSVAFPDETMPN 91421D69B57551FB csrp3-csrp3-3 1 MLP-a 3 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined MLP-b 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f5b blot overlay assays with the GST-MLP-b were performed. Western blot analysis with the GST antibody determined that the isoform does not bind to MyoD or Myogenin (Fig. 5B), although it can directly bind MLP, as detected by its interaction with MBP-MLP-a recombinant protein (Fig. 5B). In a parallel set of blot overlay assays we confirmed previous reports on the direct interaction of full length MLP with MyoD and Myogenin (Fig. 5C). csrp3-myod1 1 MyoD 5 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined MLP-a 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f5c myod1-csrp3 1 MLP-a 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Myogenin 8 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f5c csrp3-actn2 2 MLP 3 predetermined predetermined participant predetermined western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined a-actinin 4 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level T-Cap 7 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association f6c In parallel, we performed pull down assays using GST-MLP-a, which served as a positive control for these experiments (Fig. 6B). Western blot analysis of the pull down samples determined that MLP-b can bind both to α-actinin and Tcap (Fig. 6C). csrp3-csrp3-1 1 MLP-b 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined MLP inter-LIM 3 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f6e csrp3-csrp3-2 1 CFL2 6 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined MLP-a 3 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f6g cfl2-csrp3 1 MLP-a 3 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined CFL2 6 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f6h f6i we performed competition assays by blot overlay using His-CFL2 and MBP-MLP-a in the presence or absence of the GST-MLP-b. The MLP/CFL2 interaction remained unaffected by the presence of the MLP-b, indicating that the latter does not compete with CFL2 for binding to full length MLP (Fig. 6H-I).