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/ Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). Biochemical and biophysical research communications 2010 imex curation von Marschall Z., Fisher LW. lfisher@dir.nidcr.nih.gov 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro cleavage cleavage assay cleavage edman degradation edman degradation von Marschall Z., Fisher LW. I have no idea what "cABCase" might be I have added the resultant cleavage as in OLS=span the cleavage region. No appropriate feature type in menu. lfisher@dir.nidcr.nih.gov Accepted 2016-SEP-02 AT 13:22 BST AT 13:22 BST by ORCHARD Biochemical and biophysical research communications 2010 Only protein-protein interactions imex curation imex curation Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). Biochemical and biophysical research communications 2010 imex curation von Marschall Z., Fisher LW. lfisher@dir.nidcr.nih.gov 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay western blot western blot Immuno blot Immuno blot von Marschall Z., Fisher LW. I have not entered Human Decorin as isoform 1 P07585-1, as the reference quoted (pmid:3484330) is several (unclear) links and many years away (1986)- thought it better to be ambiguous rather than enter something incorrect. Proteins were purified from furin-inhibited cell culture media. Glycosaminoglycan and N-linked oligosaccharide chains were removed prior to electrophoresis. "Classic" BMP1 (BMP1–1) as used in figure 2/3 from R+D Systems, corresponds to Uniprot ID =P13497-2; a protein of 730 amino acids. Human isoform "BMP1–5" which lacks the CUB3 domain, corresponds to Uniprot ID P13497-4. The construct mDCN-4 (mouse BGN) is not mentioned in Ref 16 but only has one isoform listed in Uniprot. mTLD (mouse BMP1) clone ID6849066 -Invitrogen cannot be found, however there is also only one isoform listed in UniProt. Human Decorin, clone P2 (ref 16, Table 1) is probably a molecule of 359 amino acids -see pmid:3484330, obtained via Ref 16. lfisher@dir.nidcr.nih.gov Accepted 2016-SEP-02 AT 13:22 BST AT 13:22 BST by ORCHARD Biochemical and biophysical research communications 2010 Only protein-protein interactions imex curation imex curation bmp1_mouse Bone morphogenetic protein 1 Bmp1 Mammalian tolloid protein Procollagen C-proteinase protein protein mouse Mus musculus Mouse MPGVARPPLPLLSLPLLLLLLLLPRAGRPLDLADYTYDLGEEDAPELLNYKDPCKAAAFLGDIALDEEDLRAFQVQQAAVLRQQTARRPSIKAAGNSSALGGQGTSGQPQRESRGRWRGRPRSRRAATSRPERVWPDGVIPFVIGGNFTGSQRAVFRQAMRHWEKHTCVTFLERTDEDSYIVFTYRPCGCCSYVGRRGGGPQAISIGKNCDKFGIVVHELGHVIGFWHEHTRPDRDRHVSIVRENIQPGQEYNFLKMEVQEVESLGETYDFDSIMHYARNTFSRGIFLDTIVPKYEVNGVKPSIGQRTRLSKGDIAQARKLYKCPACGETLQDSTGNFSSPEYPNGYSAHMHCVWRISVTPGEKIILNFTSMDLYRSRLCWYDYVEVRDGFWRKAPLRGRFCGGKLPEPIVSTDSRLWVEFRSSSNWVGKGFFAVYEAICGGDVKKDNGHIQSPNYPDDYRPSKVCIWRIQVSEGFHVGLTFQSFEIERHDSCAYDYLEVRDGHSESSNLIGRYCGYEKPDDIKSTSSRLWLKFVSDGSINKAGFAVNFFKEVDECSRPNRGGCEQRCLNTLGSYKCSCDPGYELAPDKRRCEAACGGFLTKLNGSITSPGWPKEYPPNKNCIWQLVAPTQYRISLQFDFFETEGNDVCKYDFVEVRSGLTADSKLHGKFCGSEKPEVITSQYNNMRVEFKSDNTVSKKGFKAHFFSDKDECSKDNGGCQQDCVNTFGSYECQCRSGFVLHDNKHDCKEAGCEHKVTSTSGTITSPNWPDKYPSKKECTWAISSTPGHRVKLTFVEMDIESQPECAYDHLEVFDGRDAKAPVLGRFCGSKKPEPVLATGNRMFLRFYSDNSVQRKGFQASHSTECGGQVRADVKTKDLYSHAQFGDNNYPGGVDCEWVIVAEEGYGVELVFQTFEVEEETDCGYDYIELFDGYDSTAPRLGRYCGSGPPEEVYSAGDSVLVKFHSDDTISKKGFHLRYTSTKFQDTLHSRK F838441CF1DA9F1A p13497-4 Bone morphogenetic protein 1 Mammalian tolloid protein Procollagen C-proteinase BMP1 PCOLC protein protein human Homo sapiens Human MPGVARLPLLLGLLLLPRPGRPLDLADYTYDLAEEDDSEPLNYKDPCKAAAFLGDIALDEEDLRAFQVQQAVDLRRHTARKSSIKAAVPGNTSTPSCQSTNGQPQRGACGRWRGRSRSRRAATSRPERVWPDGVIPFVIGGNFTGSQRAVFRQAMRHWEKHTCVTFLERTDEDSYIVFTYRPCGCCSYVGRRGGGPQAISIGKNCDKFGIVVHELGHVVGFWHEHTRPDRDRHVSIVRENIQPGQEYNFLKMEPQEVESLGETYDFDSIMHYARNTFSRGIFLDTIVPKYEVNGVKPPIGQRTRLSKGDIAQARKLYKCPACGETLQDSTGNFSSPEYPNGYSAHMHCVWRISVTPGEKIILNFTSLDLYRSRLCWYDYVEVRDGFWRKAPLRGRFCGSKLPEPIVSTDSRLWVEFRSSSNWVGKGFFAVYEAICGGDVKKDYGHIQSPNYPDDYRPSKVCIWRIQVSEGFHVGLTFQSFEIERHDSCAYDYLEVRDGHSESSTLIGRYCGYEKPDDIKSTSSRLWLKFVSDGSINKAGFAVNFFKEVDECSRPNRGGCEQRCLNTLGSYKCSCDPGYELAPDKRRCEGCYDLQVGKPLLWDRHCFRLSTHGPEMLGTALRG May be produced at very low levels due to a premature stop codon in the mRNA, leading to nonsense-mediated mRNA decay. A47CE9FD49D5A8D7 pgs1_mouse Biglycan Bgn Bone/cartilage proteoglycan I PG-S1 protein protein mouse Mus musculus Mouse MCPLWLLTLLLALSQALPFEQKGFWDFTLDDGLLMMNDEEASGSDTTSGVPDLDSVTPTFSAMCPFGCHCHLRVVQCSDLGLKTVPKEISPDTTLLDLQNNDISELRKDDFKGLQHLYALVLVNNKISKIHEKAFSPLRKLQKLYISKNHLVEIPPNLPSSLVELRIHDNRIRKVPKGVFSGLRNMNCIEMGGNPLENSGFEPGAFDGLKLNYLRISEAKLTGIPKDLPETLNELHLDHNKIQAIELEDLLRYSKLYRLGLGHNQIRMIENGSLSFLPTLRELHLDNNKLSRVPAGLPDLKLLQVVYLHSNNITKVGINDFCPMGFGVKRAYYNGISLFNNPVPYWEVQPATFRCVTDRLAIQFGNYKK 4B57FCC9A1026BE6 pgs2_human Decorin DCN SLRR1B Bone proteoglycan II PG-S2 PG40 protein protein human Homo sapiens Human MKATIILLLLAQVSWAGPFQQRGLFDFMLEDEASGIGPEVPDDRDFEPSLGPVCPFRCQCHLRVVQCSDLGLDKVPKDLPPDTTLLDLQNNKITEIKDGDFKNLKNLHALILVNNKISKVSPGAFTPLVKLERLYLSKNQLKELPEKMPKTLQELRAHENEITKVRKVTFNGLNQMIVIELGTNPLKSSGIENGAFQGMKKLSYIRIADTNITSIPQGLPPSLTELHLDGNKISRVDAASLKGLNNLAKLGLSFNSISAVDNGSLANTPHLRELHLDNNKLTRVPGGLAEHKYIQVVYLHNNNISVVGSSDFCPPGHNTKKASYSGVSLFSNPVQYWEIQPSTFRCVYVRSAIQLGNYK FF511E871A1A52DD p13497-2 Bone morphogenetic protein 1 BMP1 PCOLC Mammalian tolloid protein Procollagen C-proteinase protein protein human Homo sapiens Human MPGVARLPLLLGLLLLPRPGRPLDLADYTYDLAEEDDSEPLNYKDPCKAAAFLGDIALDEEDLRAFQVQQAVDLRRHTARKSSIKAAVPGNTSTPSCQSTNGQPQRGACGRWRGRSRSRRAATSRPERVWPDGVIPFVIGGNFTGSQRAVFRQAMRHWEKHTCVTFLERTDEDSYIVFTYRPCGCCSYVGRRGGGPQAISIGKNCDKFGIVVHELGHVVGFWHEHTRPDRDRHVSIVRENIQPGQEYNFLKMEPQEVESLGETYDFDSIMHYARNTFSRGIFLDTIVPKYEVNGVKPPIGQRTRLSKGDIAQARKLYKCPACGETLQDSTGNFSSPEYPNGYSAHMHCVWRISVTPGEKIILNFTSLDLYRSRLCWYDYVEVRDGFWRKAPLRGRFCGSKLPEPIVSTDSRLWVEFRSSSNWVGKGFFAVYEAICGGDVKKDYGHIQSPNYPDDYRPSKVCIWRIQVSEGFHVGLTFQSFEIERHDSCAYDYLEVRDGHSESSTLIGRYCGYEKPDDIKSTSSRLWLKFVSDGSINKAGFAVNFFKEVDECSRPNRGGCEQRCLNTLGSYKCSCDPGYELAPDKRRCEAACGGFLTKLNGSITSPGWPKEYPPNKNCIWQLVAPTQYRISLQFDFFETEGNDVCKYDFVEVRSGLTADSKLHGKFCGSEKPEVITSQYNNMRVEFKSDNTVSKKGFKAHFFSEKRPALQPPRGRPHQLKFRVQKRNRTPQ DIP protein P13497-2 original sequence version: 130 DIP protein P13497-2 original sequence version: 142 E506D63729A9E86D bmp1-dcn-1 2 6 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells 7 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc protein cleavage protein cleavage 2A, 3A Human Decorin:I cannot tell from references given (Ref 16 Table 1) and backwards -Fisher 1989b- and further) if the signal sequence is still included or not.- the term "pro" rather than "pre-pro" usually means it is not. Clone P2 is the relevant construct. However from pmid:3484330 (PG-40 P2) I think the signal IS included. Use of Furin would imply it has not been cleaved? However Text states that:- "N-terminal protein sequencing remains the gold standard. The amino-terminus of the proDCN was GPFQQ.." - so this means there is NO signal sequence present. So have left range as "certain". bmp1-dcn-2 2 5 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells 7 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc protein cleavage protein cleavage 2B, 3B bmp1-dcn-3 2 4 enzyme enzyme neutral component neutral component human-bmsc human Human bmsc Bone Marrow Stromal cells 6 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells protein cleavage protein cleavage 3A stated in Methods:-"Recombinant BMP1–5 and mTLD proteins were obtained from the conditioned serum-free medium of transduced hBMSC." bmp1-dcn-4 2 3 enzyme enzyme neutral component neutral component human-bmsc human Human bmsc Bone Marrow Stromal cells 6 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells protein cleavage protein cleavage 3A-2 stated in Methods:-"Recombinant BMP1–5 and mTLD proteins were obtained from the conditioned serum-free medium of transduced hBMSC." bgn-bmp1-1 2 5 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells 4 enzyme enzyme neutral component neutral component human-bmsc human Human bmsc Bone Marrow Stromal cells protein cleavage protein cleavage 3B bgn-bmp1-2 2 5 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-bmsc human Human bmsc Bone Marrow Stromal cells 3 enzyme enzyme neutral component neutral component human-bmsc human Human bmsc Bone Marrow Stromal cells protein cleavage protein cleavage 3B-2 bmp1-dcn-5 1 6 enzyme target enzyme target substrate substrate neutral component neutral component proprotein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain glu30 biological feature biological feature certain certain sequence position certain certain certain sequence position certain resulting-cleavage resulting-cleavage human-bmsc human Human bmsc Bone Marrow Stromal cells 7 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc protein cleavage protein cleavage no figure