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 UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. FEBS J. (1742-464X) 2012 Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. in vitro In vitro protein array protein array experimental particp experimental participant identification experimental particp Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. FEBS J. (1742-464X) 2012 UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. FEBS J. (1742-464X) 2012 Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. mouse-b35 Mus musculus b35 B35 neuroblastoma cells fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. FEBS J. (1742-464X) 2012 UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. UCHL1 regulates ubiquitination and recycling of the neural cell adhesion molecule NCAM. FEBS J. (1742-464X) 2012 Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. mouse-cerebral_cortical_neuron Mouse cerebral cortical neuron brain-cortex_neuron Cerebral cortical neuron fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Wobst H., Förster S., Laurini C., Sekulla A., Dreiseidler M., Höhfeld J., Schmitz B., Diestel S. FEBS J. (1742-464X) 2012 p13591-2 Neural cell adhesion molecule 1 NCAM1 NCAM protein protein human Homo sapiens Human MLQTKDLIWTLFFLGTAVSLQVDIVPSQGEISVGESKFFLCQVAGDAKDKDISWFSPNGEKLTPNQQRISVVWNDDSSSTLTIYNANIDDAGIYKCVVTGEDGSESEATVNVKIFQKLMFKNAPTPQEFREGEDAVIVCDVVSSLPPTIIWKHKGRDVILKKDVRFIVLSNNYLQIRGIKKTDEGTYRCEGRILARGEINFKDIQVIVNVPPTIQARQNIVNATANLGQSVTLVCDAEGFPEPTMSWTKDGEQIEQEEDDEKYIFSDDSSQLTIKKVDKNDEAEYICIAENKAGEQDATIHLKVFAKPKITYVENQTAMELEEQVTLTCEASGDPIPSITWRTSTRNISSEEKASWTRPEKQETLDGHMVVRSHARVSSLTLKSIQYTDAGEYICTASNTIGQDSQSMYLEVQYAPKLQGPVAVYTWEGNQVNITCEVFAYPSATISWFRDGQLLPSSNYSNIKIYNTPSASYLEVTPDSENDFGNYNCTAVNRIGQESLEFILVQADTPSSPSIDQVEPYSSTAQVQFDEPEATGGVPILKYKAEWRAVGEEVWHSKWYDAKEASMEGIVTIVGLKPETTYAVRLAALNGKGLGEISAASEFKTQPVQGEPSAPKLEGQMGEDGNSIKVNLIKQDDGGSPIRHYLVRYRALSSEWKPEIRLPSGSDHVMLKSLDWNAEYEVYVVAENQQGKSKAAHFVFRTSAQPTAIPANGSPTSGLSTGAIVGILIVIFVLLLVVVDITCYFLNKCGLFMCIAVNLCGKAGPGAKGKDMEEGKAAFSKDESKEPIVEVRTEEERTPNHDGGKHTEPNETTPLTEPEKGPVEAKPECQETETKPAPAEVKTVPNDATQTKENESKA mint FD3B9DE80D802554 p13591-1 Neural cell adhesion molecule 1 NCAM1 NCAM N-CAM 140 protein protein human Homo sapiens Human MLQTKDLIWTLFFLGTAVSLQVDIVPSQGEISVGESKFFLCQVAGDAKDKDISWFSPNGEKLTPNQQRISVVWNDDSSSTLTIYNANIDDAGIYKCVVTGEDGSESEATVNVKIFQKLMFKNAPTPQEFREGEDAVIVCDVVSSLPPTIIWKHKGRDVILKKDVRFIVLSNNYLQIRGIKKTDEGTYRCEGRILARGEINFKDIQVIVNVPPTIQARQNIVNATANLGQSVTLVCDAEGFPEPTMSWTKDGEQIEQEEDDEKYIFSDDSSQLTIKKVDKNDEAEYICIAENKAGEQDATIHLKVFAKPKITYVENQTAMELEEQVTLTCEASGDPIPSITWRTSTRNISSEEKTLDGHMVVRSHARVSSLTLKSIQYTDAGEYICTASNTIGQDSQSMYLEVQYAPKLQGPVAVYTWEGNQVNITCEVFAYPSATISWFRDGQLLPSSNYSNIKIYNTPSASYLEVTPDSENDFGNYNCTAVNRIGQESLEFILVQADTPSSPSIDQVEPYSSTAQVQFDEPEATGGVPILKYKAEWRAVGEEVWHSKWYDAKEASMEGIVTIVGLKPETTYAVRLAALNGKGLGEISAASEFKTQPVQGEPSAPKLEGQMGEDGNSIKVNLIKQDDGGSPIRHYLVRYRALSSEWKPEIRLPSGSDHVMLKSLDWNAEYEVYVVAENQQGKSKAAHFVFRTSAQPTAIPANGSPTSGLSTGAIVGILIVIFVLLLVVVDITCYFLNKCGLFMCIAVNLCGKAGPGAKGKDMEEGKAAFSKDESKEPIVEVRTEEERTPNHDGGKHTEPNETTPLTEPEKGPVEAKPECQETETKPAPAEVKTVPNDATQTKENESKA mint homomint 68D2F0C0E6C1C2AD uchl1_human Ubiquitin carboxyl-terminal hydrolase isozyme L1 Ubiquitin thioesterase L1 Neuron cytoplasmic protein 9.5 PGP 9.5 UCHL1 protein protein human Homo sapiens Human MQLKPMEINPEMLNKVLSRLGVAGQWRFVDVLGLEEESLGSVPAPACALLLLFPLTAQHENFRKKQIEELKGQEVSPKVYFMKQTIGNSCGTIGLIHAVANNQDKLGFEDGSVLKQFLSETEKMSPEDRAKCFEKNEAIQAAHDAVAQEGQCRVDDKVNFHFILFNNVDGHLYELDGRMPFPVNHGASSEDTLLKDAAKVCREFTEREQGEVRFSAVALCKAA DIP protein P09936 original sequence version: 135 C9E972AC4DA5DA8A ncam1_mouse Neural cell adhesion molecule 1 Ncam1 Ncam protein protein mouse Mus musculus Mouse MLRTKDLIWTLFFLGTAVSLQVDIVPSQGEISVGESKFFLCQVAGDAKDKDISWFSPNGEKLSPNQQRISVVWNDDDSSTLTIYNANIDDAGIYKCVVTAEDGTQSEATVNVKIFQKLMFKNAPTPQEFKEGEDAVIVCDVVSSLPPTIIWKHKGRDVILKKDVRFIVLSNNYLQIRGIKKTDEGTYRCEGRILARGEINFKDIQVIVNVPPTVQARQSIVNATANLGQSVTLVCDADGFPEPTMSWTKDGEPIENEEEDDEKHIFSDDSSELTIRNVDKNDEAEYVCIAENKAGEQDASIHLKVFAKPKITYVENQTAMELEEQVTLTCEASGDPIPSITWRTSTRNISSEEKTLDGHMVVRSHARVSSLTLKSIQYTDAGEYICTASNTIGQDSQSMYLEFQYAPKLQGPVAVYTWEGNQVNITCEVFAYPSATISWFRDGQLLPSSNYSNIKIYNTPSASYLEVTPDSENDFGNYNCTAVNRIGQESLEFILVQADTPSSPSIDRVEPYSSTAQVQFDEPEATGGVPILKYKAEWKSLGEESWHFKWYDAKEANMEGIVTIMGLKPETRYSVRLAALNGKGLGEISAATEFKTQPVREPSAPKLEGQMGEDGNSIKVNLIKQDDGGSPIRHYLVKYRALASEWKPEIRLPSGSDHVMLKSLDWNAEYEVYVVAENQQGKSKAAHFVFRTSAQPTAIPANGSPTAGLSTGAIVGILIVIFVLLLVVMDITCYFLNKCGLLMCIAVNLCGKAGPGAKGKDMEEGKAAFSKDESKEPIVEVRTEEERTPNHDGGKHTEPNETTPLTEPELPADTTATVEDMLPSVTTVTTNSDTITETFATAQNSPTSETTTLTSSIAPPATTVPDSNSVPAGQATPSKGVTASSSSPASAPKVAPLVDLSDTPTSAPSASNLSSTVLANQGAVLSPSTPASAGETSKAPPASKASPAPTPTPAGAASPLAAVAAPATDAPQAKQEAPSTKGPDPEPTQPGTVKNPPEAATAPASPKSKAATTNPSQGEDLKMDEGNFKTPDIDLAKDVFAALGSPRPATGASGQASELAPSPADSAVPPAPAKTEKGPVETKSEPPESEAKPAPTEVKTVPNDATQTKENESKA 78AF831BABD23918 uchl1_mouse Ubiquitin carboxyl-terminal hydrolase isozyme L1 Uchl1 Ubiquitin thioesterase L1 Neuron cytoplasmic protein 9.5 PGP 9.5 protein protein mouse Mus musculus Mouse MQLKPMEINPEMLNKVLAKLGVAGQWRFADVLGLEEETLGSVPSPACALLLLFPLTAQHENFRKKQIEELKGQEVSPKVYFMKQTIGNSCGTIGLIHAVANNQDKLEFEDGSVLKQFLSETEKLSPEDRAKCFEKNEAIQAAHDSVAQEGQCRVDDKVNFHFILFNNVDGHLYELDGRMPFPVNHGASSEDSLLQDAAKVCREFTEREQGEVRFSAVALCKAA F1402BF7B0C077EA ncam1_human Neural cell adhesion molecule 1 NCAM1 NCAM protein protein human Homo sapiens Human MLQTKDLIWTLFFLGTAVSLQVDIVPSQGEISVGESKFFLCQVAGDAKDKDISWFSPNGEKLTPNQQRISVVWNDDSSSTLTIYNANIDDAGIYKCVVTGEDGSESEATVNVKIFQKLMFKNAPTPQEFREGEDAVIVCDVVSSLPPTIIWKHKGRDVILKKDVRFIVLSNNYLQIRGIKKTDEGTYRCEGRILARGEINFKDIQVIVNVPPTIQARQNIVNATANLGQSVTLVCDAEGFPEPTMSWTKDGEQIEQEEDDEKYIFSDDSSQLTIKKVDKNDEAEYICIAENKAGEQDATIHLKVFAKPKITYVENQTAMELEEQVTLTCEASGDPIPSITWRTSTRNISSEEKASWTRPEKQETLDGHMVVRSHARVSSLTLKSIQYTDAGEYICTASNTIGQDSQSMYLEVQYAPKLQGPVAVYTWEGNQVNITCEVFAYPSATISWFRDGQLLPSSNYSNIKIYNTPSASYLEVTPDSENDFGNYNCTAVNRIGQESLEFILVQADTPSSPSIDQVEPYSSTAQVQFDEPEATGGVPILKYKAEWRAVGEEVWHSKWYDAKEASMEGIVTIVGLKPETTYAVRLAALNGKGLGEISAASEFKTQPVQGEPSAPKLEGQMGEDGNSIKVNLIKQDDGGSPIRHYLVRYRALSSEWKPEIRLPSGSDHVMLKSLDWNAEYEVYVVAENQQGKSKAAHFVFRTSAQPTAIPANGSPTSGLSTGAIVGILIVIFVLLLVVVDITCYFLNKCGLFMCIAVNLCGKAGPGAKGKDMEEGKAAFSKDESKEPIVEVRTEEERTPNHDGGKHTEPNETTPLTEPEKGPVEAKPECQETETKPAPAEVKTVPNDATQTKENESKA DIP protein P13591 original sequence version: 136 FD3B9DE80D802554 ncam1-uchl1 1 UCHL1 6 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey 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 NCAM1 9 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction homomint mint To identify cytoplasmic proteins that interact with the cytoplasmic domain of human NCAM180 (hNCAM180cyt) recombinant hNCAM180cyt was expressed and purified by ligand affinity chromatography (Figure 1). Purified hNCAM180cyt was applied on a protein macroarray comprising 25300 expression clones of human fetal brain to screen for hNCAM180cyt binding partners. Detailed analysis of expression clones which had bound hNCAM180cyt revealed already identified binding partners of NCAM, i.e. PLCγ [15], spectrin [9,10], tubulin [14] and PP1 [15]. Additionally, we detected further proteins that have not yet been identified as interaction partners of NCAM s cytoplasmic domain, interestingly including UCHL1 as a potential interaction partner of hNCAM180cyt. n f ncam1-uchl1-1 3 Uchl1 8 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell NCAM1 7 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f2a mint Endogenous NCAM and UCHL1 colocalized partially in cell somata and neurites of primary cortical neurons (Figure 2A) ncam1-uchl1-2 2 NCAM180 4 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell Uchl1 8 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization mint NCAM180 or NCAM140, respectively, was expressed in B35 cells and analyzed for colocalization with endogenous UCHL1. For both NCAM isoforms a partial colocalization could also be observed in B35 cells (Figure 2B). If UCHL1 plays a role in recycling of NCAM to the cell surface it has to be associated not only with cell surface NCAM but particularly with internalized NCAM. Therefore we analyzed whether internalized NCAM colocalizes with UCHL1. In B35 cells, expressing either NCAM180 or NCAM140, a small portion of internalized NCAM colocalized with UCHL1 f2b f5a f5b f5c ncam1-uchl1-3 2 NCAM140 5 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell Uchl1 8 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization mint NCAM180 or NCAM140, respectively, was expressed in B35 cells and analyzed for colocalization with endogenous UCHL1. For both NCAM isoforms a partial colocalization could also be observed in B35 cells (Figure 2B). If UCHL1 plays a role in recycling of NCAM to the cell surface it has to be associated not only with cell surface NCAM but particularly with internalized NCAM. Therefore we analyzed whether internalized NCAM colocalizes with UCHL1. In B35 cells, expressing either NCAM180 or NCAM140, a small portion of internalized NCAM colocalized with UCHL1 f2b f5d f5e f5f