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 Effect of Ca2+ on the microtubule-severing enzyme p60-katanin. Insight into the substrate-dependent activation mechanism. Effect of Ca2+ on the microtubule-severing enzyme p60-katanin. Insight into the substrate-dependent activation mechanism. FEBS J. (1742-464X) 2012 Iwaya N., Akiyama K., Goda N., Tenno T., Fujiwara Y., Hamada D., Ikura T., Shirakawa M., Hiroaki H. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Iwaya N., Akiyama K., Goda N., Tenno T., Fujiwara Y., Hamada D., Ikura T., Shirakawa M., Hiroaki H. FEBS J. (1742-464X) 2012 Effect of Ca2+ on the microtubule-severing enzyme p60-katanin. Insight into the substrate-dependent activation mechanism. Effect of Ca2+ on the microtubule-severing enzyme p60-katanin. Insight into the substrate-dependent activation mechanism. FEBS J. (1742-464X) 2012 Iwaya N., Akiyama K., Goda N., Tenno T., Fujiwara Y., Hamada D., Ikura T., Shirakawa M., Hiroaki H. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Iwaya N., Akiyama K., Goda N., Tenno T., Fujiwara Y., Hamada D., Ikura T., Shirakawa M., Hiroaki H. FEBS J. (1742-464X) 2012 ktna1_mouse Katanin p60 ATPase-containing subunit A1 Katna1 Lipotransin p60 katanin protein protein mouse Mus musculus Mouse MSLQMIVENVKLAREYALLGNYDSAMVYYQGVLDQMNKYLYSVKDTHLRQKWQQVWQEINVEAKQVKDIMKTLESFKLDITSLQAAQHELPAAEGEVWSLPVPVERRPLPGPRKRQSSQHSDPKPHSNRPSTVVRAHRPSPQNLHNDRGKAVRSREKKEQSKGREEKNKLPAAVTEPEANKFDGTGYDKDLVEALERDIISQNPNVRWYDIADLVEAKKLLQEAVVLPMWMPEFFKGIRRPWKGVLMVGPPGTGKTLLAKAVATECKTTFFNVSSSTLTSKYRGESEKLVRLLFEMARFYSPATIFIDEIDSICSRRGTSEEHEASRRMKAELLVQMDGVGGASENDDPSKMVMVLAATNFPWDIDEALRRRLEKRIYIPLPSAKGREELLRISLRELELADDVNLASIAENMEGYSGADITNVCRDASLMAMRRRIEGLTPEEIRNLSREAMHMPTTMEDFEMALKKISKSVSAADIERYEKWIVEFGSC mint 3F02C73FDE94DE42 ktnb1_mouse Katanin p80 WD40 repeat-containing subunit B1 p80 katanin Katnb1 protein protein mouse Mus musculus Mouse MATPVVTKTAWKLQEIVAHASNVSSLVLGKASGRLLATGGDDCRVNLWSINKPNCIMSLTGHTSPVESVRLNTPEELIVAGSQSGSIRVWDLEAAKILRTLMGHKANICSLDFHPYGEFVASGSQDTNIKLWDIRRKGCVFRYRGHSQAVRCLRFSPDGKWLASAADDHTVKLWDLTAGKMMSEFPGHTGPVNVVEFHPNEYLLASGSSDRTIRFWDLEKFQVVSCIEGEPGPVRSVLFNPDGCCLYSGCQDSLRVYGWEPERCFDVVLVNWGKVADLAICNDQLIGVAFSQSNVSSYVVDLTRVTRTGTVTQDPVQANQPLTQQTPNPGVSLRRIYERPSTTCSKPQRVKHNSESERRSPSSEDDRDERESRAEIQNAEDYNEIFQPKNSISRTPPRRSEPFPAPPEDDAATVKEVSKPSPAMDVQLPQLPVPNLEVPARPSVMTSTPAPKGEPDIIPATRNEPIGLKASDFLPAVKVPQQAELVDEDAMSQIRKGHDTMFVVLTSRHKNLDTVRAVWTTGDIKTSVDSAVAINDLSVVVDLLNIVNQKASLWKLDLCTTVLPQIEKLLQSKYESYVQTGCTSLKLILQRFLPLITDILAAPPSVGVDISREERLHKCRLCFKQLKSISGLVKSKSGLSGRHGSAFRELHLLMASLD mint Participates in a complex which severs microtubules in an ATP-dependent manner. May act to target the enzymatic subunit of this complex to sites of action such as the centrosome. Microtubule severing may promote rapid reorganization of cellular microtubule arrays and the release of microtubules from the centrosome following nucleation. Microtubule release from the mitotic spindle poles may allow depolymerization of the microtubule end proximal to the spindle pole, leading to poleward microtubule flux and poleward motion of chromosome. Microtubule release within the cell body of neurons may be required for their transport into neuronal processes by microtubule-dependent motor proteins. This transport is required for axonal growth CB04B82DA34A19EF katna1-katnb1 1 p80-CTD 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain p60-vMIT 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction mint Next, we determined the interface between p60-vMIT and p80-CTD by 1H-15N HSQC experiments. The binding of p80-CTD induced drastic line broadening of NH signals from 15N-labeled p60-vMIT (Fig. 2A; Fig. S1A). Mapping of the broadened signals upon binding to p80-CTD suggested that a wide area covering the surface of helices 1/2 and helices 1/3 of p60-vMIT might form the binding site for p80-CTD (Fig. 2B; Fig. S1B; Table S1). f2a f2b sf1a sf1b st1 katnb1-katna1 2 p60-vMIT 3 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain p80-CTD 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2c mint We further examined the key residues involved in p80-binding by using Ala-substituted mutants of p60-vMIT in an in vitro pull-down assay (Fig. 2C). Wild-type p60-vMIT bound to p80-CTD in nearly a 1 to 1 ratio, as judged from the density of the bands in SDS-PAGE (Fig. 2C, lanes 3 and 14). All Ala mutants also showed similar binding to p80-CTD. Therefore, we assumed that a polar single mutation does not completely inhibit the interaction between p60-vMIT and p80-CTD, probably because the interface is multivalent and covers a wide area including helix 1.