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 http://mint.bio.uniroma2.it/mint thomas-2013-1 DELINEATION OF THE TRAK BINDING REGIONS OF THE KINESIN-1 MOTOR PROTEINS human-293a Homo sapiens hek293a hek293a anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Randall T., Moores C., Anne Stephenson F. anne.stephenson@ucl.ac.uk FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation trak2_rat Trafficking kinesin-binding protein 2 Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 3 protein homolog GABA-A receptor-interacting factor 1 O-GlcNAc transferase-interacting protein of 98 kDa Trak2 Als2cr3 Grif1 Oip98 protein protein rat Rattus norvegicus (Rat) Rat MSLSQNAIFKSQTGEENLMSSNHRDSESITDVCSNEDLPEVELVNLLEEQLPQYKLRVDSLFLYENQDWSQSSHQQQDASETLSPVLAEETFRYMILGTDRVEQMTKTYNDIDMVTHLLAERDRDLELAARIGQALLKRNHVLSEQNESLEEQLGQAFDQVNQLQHELSKKEELLRIVSIASEESETDSSCSTPLRFNESFSLSQGLLQLDMMHEKLKELEEENMALRSKACHIKTETFTYEEKEQKLINDCVNELRETNAQMSRMTEELSGKSDELLRYQEEISSLLSQIVDLQHKLKEHVIEKEELRLHLQASKDAQRQLTMELHELQDRNMECLGMLHESQEEIKELRNKAGPSAHLCFSQAYGVFAGESLAAEIEGTMRKKLSLDEESVFKQKAQQKRVFDTVKVANDTRGRSVTFPVLLPIPGSNRSSVIMTAKPFESGVQQTEDKTLPNQGSSTEVPGNSHPRDPPGLPEDSDLATALHRLSLRRQNYLSEKQFFAEEWERKLQILAEQEEEVSSCEALTENLASFCTDQSETTELGSAGCLRGFMPEKLQIVKPLEGSQTLHHWQQLAQPNLGTILDPRPGVITKGFTQMPKDAVYHISDLEEDEEVGITFQVQQPLQLEQKPAPPPPVTGIFLPPMTSAGGPVSVATSNPGKCLSFTNSTFTFTTCRILHPSDITQVTPSSGFPSLSCGSSAGSASNTAVNSPAASYRLSIGESITNRRDSTITFSSTRSLAKLLQERGISAKVYHSPASENPLLQLRPKALATPSTPPNSPSQSPCSSPVPFEPRVHVSENFLASRPAETFLQEMYGLRPSRAPPDVGQLKMNLVDRLKRLGIARVVKTPVPRENGKSREAEMGLQKPDSAVYLNSGGSLLGGLRRNQSLPVMMGSFGAPVCTTSPKMGILKED kif5a_human Kinesin heavy chain isoform 5A Neuronal kinesin heavy chain Kinesin heavy chain neuron-specific 1 KIF5A NKHC1 protein protein human Homo sapiens Human MAETNNECSIKVLCRFRPLNQAEILRGDKFIPIFQGDDSVVIGGKPYVFDRVFPPNTTQEQVYHACAMQIVKDVLAGYNGTIFAYGQTSSGKTHTMEGKLHDPQLMGIIPRIARDIFNHIYSMDENLEFHIKVSYFEIYLDKIRDLLDVTKTNLSVHEDKNRVPFVKGCTERFVSSPEEILDVIDEGKSNRHVAVTNMNEHSSRSHSIFLINIKQENMETEQKLSGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALAEGTKSYVPYRDSKMTRILQDSLGGNCRTTMFICCSPSSYNDAETKSTLMFGQRAKTIKNTASVNLELTAEQWKKKYEKEKEKTKAQKETIAKLEAELSRWRNGENVPETERLAGEEAALGAELCEETPVNDNSSIVVRIAPEERQKYEEEIRRLYKQLDDKDDEINQQSQLIEKLKQQMLDQEELLVSTRGDNEKVQRELSHLQSENDAAKDEVKEVLQALEELAVNYDQKSQEVEEKSQQNQLLVDELSQKVATMLSLESELQRLQEVSGHQRKRIAEVLNGLMKDLSEFSVIVGNGEIKLPVEISGAIEEEFTVARLYISKIKSEVKSVVKRCRQLENLQVECHRKMEVTGRELSSCQLLISQHEAKIRSLTEYMQSVELKKRHLEESYDSLSDELAKLQAQETVHEVALKDKEPDTQDADEVKKALELQMESHREAHHRQLARLRDEINEKQKTIDELKDLNQKLQLELEKLQADYEKLKSEEHEKSTKLQELTFLYERHEQSKQDLKGLEETVARELQTLHNLRKLFVQDVTTRVKKSAEMEPEDSGGIHSQKQKISFLENNLEQLTKVHKQLVRDNADLRCELPKLEKRLRATAERVKALEGALKEAKEGAMKDKRRYQQEVDRIKEAVRYKSSGKRGHSAQIAKPVRPGHYPASSPTNPYGTRSPECISYTNSLFQNYQNLYLQATPSSTSDMYFANSCTSSGATSSGGPLASYQKANMDNGNATDINDNRSDLPCGYEAEDQAKLFPLHQETAAS kif5c_human Kinesin heavy chain isoform 5C Kinesin heavy chain neuron-specific 2 KIF5C KIAA0531 NKHC2 protein protein human Homo sapiens Human MADPAECSIKVMCRFRPLNEAEILRGDKFIPKFKGDETVVIGQGKPYVFDRVLPPNTTQEQVYNACAKQIVKDVLEGYNGTIFAYGQTSSGKTHTMEGKLHDPQLMGIIPRIAHDIFDHIYSMDENLEFHIKVSYFEIYLDKIRDLLDVSKTNLAVHEDKNRVPYVKGCTERFVSSPEEVMDVIDEGKANRHVAVTNMNEHSSRSHSIFLINIKQENVETEKKLSGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALAEGTKTHVPYRDSKMTRILQDSLGGNCRTTIVICCSPSVFNEAETKSTLMFGQRAKTIKNTVSVNLELTAEEWKKKYEKEKEKNKTLKNVIQHLEMELNRWRNGEAVPEDEQISAKDQKNLEPCDNTPIIDNIAPVVAGISTEEKEKYDEEISSLYRQLDDKDDEINQQSQLAEKLKQQMLDQDELLASTRRDYEKIQEELTRLQIENEAAKDEVKEVLQALEELAVNYDQKSQEVEDKTRANEQLTDELAQKTTTLTTTQRELSQLQELSNHQKKRATEILNLLLKDLGEIGGIIGTNDVKTLADVNGVIEEEFTMARLYISKMKSEVKSLVNRSKQLESAQMDSNRKMNASERELAACQLLISQHEAKIKSLTDYMQNMEQKRRQLEESQDSLSEELAKLRAQEKMHEVSFQDKEKEHLTRLQDAEEMKKALEQQMESHREAHQKQLSRLRDEIEEKQKIIDEIRDLNQKLQLEQEKLSSDYNKLKIEDQEREMKLEKLLLLNDKREQAREDLKGLEETVSRELQTLHNLRKLFVQDLTTRVKKSVELDNDDGGGSAAQKQKISFLENNLEQLTKVHKQLVRDNADLRCELPKLEKRLRATAERVKALESALKEAKENAMRDRKRYQQEVDRIKEAVRAKNMARRAHSAQIAKPIRPGHYPASSPTAVHAIRGGGGSSSNSTHYQK trak1_human Trafficking kinesin-binding protein 1 Protein Milton TRAK1 KIAA1042 OIP106 106 kDa O-GlcNAc transferase-interacting protein protein protein human Homo sapiens Human MALVFQFGQPVRAQPLPGLCHGKLIRTNACDVCNSTDLPEVEIISLLEEQLPHYKLRADTIYGYDHDDWLHTPLISPDANIDLTTEQIEETLKYFLLCAERVGQMTKTYNDIDAVTRLLEEKERDLELAARIGQSLLKKNKTLTERNELLEEQVEHIREEVSQLRHELSMKDELLQFYTSAAEESEPESVCSTPLKRNESSSSVQNYFHLDSLQKKLKDLEEENVVLRSEASQLKTETITYEEKEQQLVNDCVKELRDANVQIASISEELAKKTEDAARQQEEITHLLSQIVDLQKKAKACAVENEELVQHLGAAKDAQRQLTAELRELEDKYAECMEMLHEAQEELKNLRNKTMPNTTSRRYHSLGLFPMDSLAAEIEGTMRKELQLEEAESPDITHQKRVFETVRNINQVVKQRSLTPSPMNIPGSNQSSAMNSLLSSCVSTPRSSFYGSDIGNVVLDNKTNSIILETEAADLGNDERSKKPGTPGTPGSHDLETALRRLSLRRENYLSERRFFEEEQERKLQELAEKGELRSGSLTPTESIMSLGTHSRFSEFTGFSGMSFSSRSYLPEKLQIVKPLEGSATLHHWQQLAQPHLGGILDPRPGVVTKGFRTLDVDLDEVYCLNDFEEDDTGDHISLPRLATSTPVQHPETSAHHPGKCMSQTNSTFTFTTCRILHPSDELTRVTPSLNSAPTPACGSTSHLKSTPVATPCTPRRLSLAESFTNTRESTTTMSTSLGLVWLLKERGISAAVYDPQSWDRAGRGSLLHSYTPKMAVIPSTPPNSPMQTPTSSPPSFEFKCTSPPYDNFLASKPASSILREVREKNVRSSESQTDVSVSNLNLVDKVRRFGVAKVVNSGRAHVPTLTEEQGPLLCGPPGPAPALVPRGLVPEGLPLRCPTVTSAIGGLQLNSGIRRNRSFPTMVGSSMQMKAPVTLTSGILMGAKLSKQTSLR kif5a-trak2 1444187 n/a TRAK2 1444188 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a KIF5A 1444189 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed binding region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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 false physical association physical association false false false f2 f3 Thus using this approach, three regions within the KIF5A Cterminal region were shown to contribute to the TRAK2 binding site i.e. KIF5A 942-961; KIF5A 877- 885 and KIF5A 861-877. (Note that full immunoblots for Figures 3 – 5 can be viewed in the Supplementary Information, Figures S1- S3). The region KIF5A 877-883 was studied in more detail. Three additional truncations were made at KIF5A 883, KIF5A 881 and KIF5A 879 plus the region 877- 883 was deleted. The constructs truncated at KIF5A 883, KIF5A 881 and KIF5A 879 co-immunoprecipitated with TRAK2 with a similar efficiency to KIF5A1-885 as indeed did the KIF5AΔ877-883 deletion. The latter was unexpected since this region, amino acid sequence GALKEAKE, had been clearly implicated in the efficiency of KIF5A/TRAK2 association (Figure 3D). It may be explained by an alteration of the KIF5A tertiary structure such that other amino acids requisite for the binding of TRAK2 are now available to compensate for the deleted region thus the predicted loss in efficacy of co-immunoprecipitation is circumvented. In the context of the full length KIF5A therefore, the other sites of interaction are sufficient to support partial association with TRAK2 even when 877-883 is removed. trak2-kif5a 1444187 n/a TRAK2 1444188 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a KIF5A 1444189 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed binding region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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 false physical association physical association false false false f2 f3 Thus using this approach, three regions within the KIF5A Cterminal region were shown to contribute to the TRAK2 binding site i.e. KIF5A 942-961; KIF5A 877- 885 and KIF5A 861-877. (Note that full immunoblots for Figures 3 – 5 can be viewed in the Supplementary Information, Figures S1- S3). The region KIF5A 877-883 was studied in more detail. Three additional truncations were made at KIF5A 883, KIF5A 881 and KIF5A 879 plus the region 877- 883 was deleted. The constructs truncated at KIF5A 883, KIF5A 881 and KIF5A 879 co-immunoprecipitated with TRAK2 with a similar efficiency to KIF5A1-885 as indeed did the KIF5AΔ877-883 deletion. The latter was unexpected since this region, amino acid sequence GALKEAKE, had been clearly implicated in the efficiency of KIF5A/TRAK2 association (Figure 3D). It may be explained by an alteration of the KIF5A tertiary structure such that other amino acids requisite for the binding of TRAK2 are now available to compensate for the deleted region thus the predicted loss in efficacy of co-immunoprecipitation is circumvented. In the context of the full length KIF5A therefore, the other sites of interaction are sufficient to support partial association with TRAK2 even when 877-883 is removed. trak2-kif5c-2 1444187 n/a TRAK2 1444188 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a KIF5C 1444190 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false f4 Thus using this approach, three regions within the KIF5A Cterminal region were shown to contribute to the TRAK2 binding site i.e. KIF5A 942-961; KIF5A 877- 885 and KIF5A 861-877. (Note that full immunoblots for Figures 3 – 5 can be viewed in the Supplementary Information, Figures S1- S3). The region KIF5A 877-883 was studied in more detail. Three additional truncations were made at KIF5A 883, KIF5A 881 and KIF5A 879 plus the region 877- 883 was deleted. The constructs truncated at KIF5A 883, KIF5A 881 and KIF5A 879 co-immunoprecipitated with TRAK2 with a similar efficiency to KIF5A1-885 as indeed did the KIF5AΔ877-883 deletion. The latter was unexpected since this region, amino acid sequence GALKEAKE, had been clearly implicated in the efficiency of KIF5A/TRAK2 association (Figure 3D). It may be explained by an alteration of the KIF5A tertiary structure such that other amino acids requisite for the binding of TRAK2 are now available to compensate for the deleted region thus the predicted loss in efficacy of co-immunoprecipitation is circumvented. In the context of the full length KIF5A therefore, the other sites of interaction are sufficient to support partial association with TRAK2 even when 877-883 is removed. trak1-kif5a-1 1444187 n/a KIF5A 1444189 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 false binding region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n/a TRAK1 1444191 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed myv tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f5 Thus using this approach, three regions within the KIF5A Cterminal region were shown to contribute to the TRAK2 binding site i.e. KIF5A 942-961; KIF5A 877- 885 and KIF5A 861-877. (Note that full immunoblots for Figures 3 – 5 can be viewed in the Supplementary Information, Figures S1- S3). The region KIF5A 877-883 was studied in more detail. Three additional truncations were made at KIF5A 883, KIF5A 881 and KIF5A 879 plus the region 877- 883 was deleted. The constructs truncated at KIF5A 883, KIF5A 881 and KIF5A 879 co-immunoprecipitated with TRAK2 with a similar efficiency to KIF5A1-885 as indeed did the KIF5AΔ877-883 deletion. The latter was unexpected since this region, amino acid sequence GALKEAKE, had been clearly implicated in the efficiency of KIF5A/TRAK2 association (Figure 3D). It may be explained by an alteration of the KIF5A tertiary structure such that other amino acids requisite for the binding of TRAK2 are now available to compensate for the deleted region thus the predicted loss in efficacy of co-immunoprecipitation is circumvented. In the context of the full length KIF5A therefore, the other sites of interaction are sufficient to support partial association with TRAK2 even when 877-883 is removed.