IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Genomics (0888-7543) 2001 imex curation Pan H., Qin WX., Huo KK., Wan DF., Yu Y., Xu ZG., Hu QD., Gu KT., Zhou XM., Jiang HQ., Zhang PP., Huang Y., Li YY., Gu JR. Cancer - Interactions investigated in the context of cancer nlorg@public.sta.net.cn 08-DEC-2006: Contacted by JYOTI. Two cDNA libraries were used, one derived from human fetal brain and one from human liver (OriGene). Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Pan H., Qin WX., Huo KK., Wan DF., Yu Y., Xu ZG., Hu QD., Gu KT., Zhou XM., Jiang HQ., Zhang PP., Huang Y., Li YY., Gu JR. Cancer - Interactions investigated in the context of cancer nlorg@public.sta.net.cn Accepted 2006-DEC-4 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Two cDNA libraries were used, one derived from human fetal brain and one from human liver (OriGene). Genomics (0888-7543) 2001 Only protein-protein interactions imex curation Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Genomics (0888-7543) 2001 imex curation Pan H., Qin WX., Huo KK., Wan DF., Yu Y., Xu ZG., Hu QD., Gu KT., Zhou XM., Jiang HQ., Zhang PP., Huang Y., Li YY., Gu JR. Cancer - Interactions investigated in the context of cancer nlorg@public.sta.net.cn 08-DEC-2006: Contacted by JYOTI. Two cDNA libraries were used, one derived from human fetal brain and one from human liver (OriGene). Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Pan H., Qin WX., Huo KK., Wan DF., Yu Y., Xu ZG., Hu QD., Gu KT., Zhou XM., Jiang HQ., Zhang PP., Huang Y., Li YY., Gu JR. Cancer - Interactions investigated in the context of cancer nlorg@public.sta.net.cn Accepted 2006-DEC-4 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Two cDNA libraries were used, one derived from human fetal brain and one from human liver (OriGene). Genomics (0888-7543) 2001 Only protein-protein interactions imex curation asgr2_human Asialoglycoprotein receptor 2 ASGR2 Hepatic lectin H2 CLEC4H2 C-type lectin domain family 4 member H2 protein protein human Homo sapiens Human MAKDFQDIQQLSSEENDHPFHQGEGPGTRRLNPRRGNPFLKGPPPAQPLAQRLCSMVCFSLLALSFNILLLVVICVTGSQSEGHGGAQLQAELRSLKEAFSNFSSSTLTEVQAISTHGGSVGDKITSLGAKLEKQQQDLKADHDALLFHLKHFPVDLRFVACQMELLHSNGSQRTCCPVNWVEHQGSCYWFSHSGKAWAEAEKYCQLENAHLVVINSWEEQKFIVQHTNPFNTWIGLTDSDGSWKWVDGTDYRHNYKNWAVTQPDNWHGHELGGSEDCVEVQPDGRWNDDFCLQVYRWVCEKRRNATGEVA C86D2247FFF604DC s22a1_human Solute carrier family 22 member 1 SLC22A1 OCT1 Organic cation transporter 1 protein protein human Homo sapiens Human MPTVDDILEQVGESGWFQKQAFLILCLLSAAFAPICVGIVFLGFTPDHHCQSPGVAELSQRCGWSPAEELNYTVPGLGPAGEAFLGQCRRYEVDWNQSALSCVDPLASLATNRSHLPLGPCQDGWVYDTPGSSIVTEFNLVCADSWKLDLFQSCLNAGFLFGSLGVGYFADRFGRKLCLLGTVLVNAVSGVLMAFSPNYMSMLLFRLLQGLVSKGNWMAGYTLITEFVGSGSRRTVAIMYQMAFTVGLVALTGLAYALPHWRWLQLAVSLPTFLFLLYYWCVPESPRWLLSQKRNTEAIKIMDHIAQKNGKLPPADLKMLSLEEDVTEKLSPSFADLFRTPRLRKRTFILMYLWFTDSVLYQGLILHMGATSGNLYLDFLYSALVEIPGAFIALITIDRVGRIYPMAMSNLLAGAACLVMIFISPDLHWLNIIIMCVGRMGITIAIQMICLVNAELYPTFVRNLGVMVCSSLCDIGGIITPFIVFRLREVWQALPLILFAVLGLLAAGVTLLLPETKGVALPETMKDAENLGRKAKPKENTIYLKVQTSEPSGT 55206B897DE32202 asgr1_human Asialoglycoprotein receptor 1 ASGR1 Hepatic lectin H1 CLEC4H1 C-type lectin domain family 4 member H1 protein protein human Homo sapiens Human MTKEYQDLQHLDNEESDHHQLRKGPPPPQPLLQRLCSGPRLLLLSLGLSLLLLVVVCVIGSQNSQLQEELRGLRETFSNFTASTEAQVKGLSTQGGNVGRKMKSLESQLEKQQKDLSEDHSSLLLHVKQFVSDLRSLSCQMAALQGNGSERTCCPVNWVEHERSCYWFSRSGKAWADADNYCRLEDAHLVVVTSWEEQKFVQHHIGPVNTWMGLHDQNGPWKWVDGTDYETGFKNWRPEQPDDWYGHGLGGGEDCAHFTDDGRWNDDVCQRPYRWVCETELDKASQEPPLL 9B825527B2FC1FD0 cers2_human Ceramide synthase 2 CERS2 TMSG1 Tumor metastasis-suppressor gene 1 protein SP260 LASS2 LAG1 longevity assurance homolog 2 Sphingosine N-acyltransferase CERS2 Very-long-chain ceramide synthase CERS2 protein protein human Homo sapiens Human MLQTLYDYFWWERLWLPVNLTWADLEDRDGRVYAKASDLYITLPLALLFLIVRYFFELYVATPLAALLNIKEKTRLRAPPNATLEHFYLTSGKQPKQVEVELLSRQSGLSGRQVERWFRRRRNQDRPSLLKKFREASWRFTFYLIAFIAGMAVIVDKPWFYDMKKVWEGYPIQSTIPSQYWYYMIELSFYWSLLFSIASDVKRKDFKEQIIHHVATIILISFSWFANYIRAGTLIMALHDSSDYLLESAKMFNYAGWKNTCNNIFIVFAIVFIITRLVILPFWILHCTLVYPLELYPAFFGYYFFNSMMGVLQLLHIFWAYLILRMAHKFITGKLVEDERSDREETESSEGEEAAAGGGAKSRPLANGHPILNNNHRKND C8CEAF6AAD0B4577 vatl_human V-type proton ATPase 16 kDa proteolipid subunit ATP6V0C ATP6C ATP6L ATPL Vacuolar proton pump 16 kDa proteolipid subunit protein protein human Homo sapiens Human MSESKSGPEYASFFAVMGASAAMVFSALGAAYGTAKSGTGIAAMSVMRPEQIMKSIIPVVMAGIIAIYGLVVAVLIANSLNDDISLYKSFLQLGAGLSVGLSGLAAGFAIGIVGDAGVRGTAQQPRLFVGMILILIFAEVLGLYGLIVALILSTK 91141854A0492A5B lass2-asgr1 2 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Of the 43 clones, 41 passed the mating test for interaction specificity. GenBank database searches showed that 14 of the 41 clones encoded seven proteins of known function. All were membrane-associated receptors or transporters. Among them, four were confirmed to physically interact with LASS2 in the GST pull-down assay as described below: the high-affinity asialoglycoprotein receptors type 1 (AGPRH1; GenBank acc. no. M10058) and type 2 (AGPRH2; GenBank acc. no. U97179), the organic cation transporter-1 (OCT1; GenBank acc. no. U77086), and the proteolipid subunit of vacuolar H+ATPase (VPL; GenBank acc. no. M62762) The UniprotKB sequence P07306 for the asialoglycoprotein receptor type 1 does not have the initial Met found in the sequence referred to by the authors (GenBank M10058). lass2-asgr2 2 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Of the 43 clones, 41 passed the mating test for interaction specificity. GenBank database searches showed that 14 of the 41 clones encoded seven proteins of known function. All were membrane-associated receptors or transporters. Among them, four were confirmed to physically interact with LASS2 in the GST pull-down assay as described below: the high-affinity asialoglycoprotein receptors type 1 (AGPRH1; GenBank acc. no. M10058) and type 2 (AGPRH2; GenBank acc. no. U97179), the organic cation transporter-1 (OCT1; GenBank acc. no. U77086), and the proteolipid subunit of vacuolar H+ATPase (VPL; GenBank acc. no. M62762) lass2-hoct1 2 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range 6 unspecified role unspecified role bait bait physical association physical association Of the 43 clones, 41 passed the mating test for interaction specificity. GenBank database searches showed that 14 of the 41 clones encoded seven proteins of known function. All were membrane-associated receptors or transporters. Among them, four were confirmed to physically interact with LASS2 in the GST pull-down assay as described below: the high-affinity asialoglycoprotein receptors type 1 (AGPRH1; GenBank acc. no. M10058) and type 2 (AGPRH2; GenBank acc. no. U97179), the organic cation transporter-1 (OCT1; GenBank acc. no. U77086), and the proteolipid subunit of vacuolar H+ATPase (VPL; GenBank acc. no. M62762) The authors state that the 5' ends of the cDNAs for OCT1 began at +917 and +973. Thus the shortest construct begins at amino acid 325. However, they do not specify where the 3' end finishes. lass2-atp6v0c 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Of the 43 clones, 41 passed the mating test for interaction specificity. GenBank database searches showed that 14 of the 41 clones encoded seven proteins of known function. All were membrane-associated receptors or transporters. Among them, four were confirmed to physically interact with LASS2 in the GST pull-down assay as described below: the high-affinity asialoglycoprotein receptors type 1 (AGPRH1; GenBank acc. no. M10058) and type 2 (AGPRH2; GenBank acc. no. U97179), the organic cation transporter-1 (OCT1; GenBank acc. no. U77086), and the proteolipid subunit of vacuolar H+ATPase (VPL; GenBank acc. no. M62762) asgr1-lass2 1 5 unspecified role unspecified role bait bait region 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 ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction The UniprotKB sequence P07306 for the asialoglycoprotein receptor type 1 does not have the initial Met found in the sequence referred to by the authors (GenBank M10058). we used a GST pull-down assay with biotinylated in vitro translated LASS2 and GST fusion proteins of AGPRH1, AGPRH2, VPL, and the C-terminal half of OCT1 (amino acids 306-544). Confirming the interactions identified in the twohybrid assay, we found that LASS2 bound to GST-AGPRH1, GST-AGPRH2, GST-VPL, and GST-OCT1 (amino acids 306-504), but not to GST alone 4 asgr2-lass2 1 3 unspecified role unspecified role bait bait region 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 ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction we used a GST pull-down assay with biotinylated in vitro translated LASS2 and GST fusion proteins of AGPRH1, AGPRH2, VPL, and the C-terminal half of OCT1 (amino acids 306-544). Confirming the interactions identified in the twohybrid assay, we found that LASS2 bound to GST-AGPRH1, GST-AGPRH2, GST-VPL, and GST-OCT1 (amino acids 306-504), but not to GST alone 4 hoct1-lass2 1 6 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 4 unspecified role unspecified role bait bait region 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction we used a GST pull-down assay with biotinylated in vitro translated LASS2 and GST fusion proteins of AGPRH1, AGPRH2, VPL, and the C-terminal half of OCT1 (amino acids 306-544). Confirming the interactions identified in the twohybrid assay, we found that LASS2 bound to GST-AGPRH1, GST-AGPRH2, GST-VPL, and GST-OCT1 (amino acids 306-504), but not to GST alone 4 It is specified that the hOCT1 construct begins at residue 306 and is the C-terminal half of the protein. However, the authors state that the construct ends at residue 544 and also that it ends at 504. The translation of the GenBank sequence they refer to (U77086)is 554 residues long as the UniprotKB sequence. hoct1-lass2-1 1 7 unspecified role unspecified role bait bait region 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 ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction we used a GST pull-down assay with biotinylated in vitro translated LASS2 and GST fusion proteins of AGPRH1, AGPRH2, VPL, and the C-terminal half of OCT1 (amino acids 306-544). Confirming the interactions identified in the twohybrid assay, we found that LASS2 bound to GST-AGPRH1, GST-AGPRH2, GST-VPL, and GST-OCT1 (amino acids 306-504), but not to GST alone 4