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/ Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. J. Biol. Chem. (0021-9258) 2014 imex curation Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Only protein-protein interactions in vitro In vitro protease assay protease assay western blot western blot Immuno blot Immuno blot Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs and hence results obtained from their use are of dubious biological relevence. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Accepted 2015-FEB-27 by ORCHARD J. Biol. Chem. (0021-9258) 2014 Only protein-protein interactions imex curation Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. J. Biol. Chem. (0021-9258) 2014 imex curation Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Only protein-protein interactions human-skin Homo sapiens skin skin pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. I have requested an update for Caspase 14 from UniProt as the last reference quoted is 2004. (sequences of cleaved chains in UniProt are not currently the same as those given by authors) normal human skin An antibody (h14D146) was used which recogises the "active" form of Caspase -14: the antibody reacts with the large subunit -which is stated by the authors to be amino acids 6-146, but the antibody does not react with the pro-form. (see reference 4.) Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Accepted 2015-FEB-27 by ORCHARD J. Biol. Chem. (0021-9258) 2014 Only protein-protein interactions imex curation Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. J. Biol. Chem. (0021-9258) 2014 imex curation Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Only protein-protein interactions in vitro In vitro pull down pull down mudpit multidimensional protein identification technology mudpit Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Human keratinocytes were derived from normal foreskin. Cornified cell extracts (skin) were prepared from scraped heels of healthy individuals. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Accepted 2015-FEB-27 by ORCHARD J. Biol. Chem. (0021-9258) 2014 Only protein-protein interactions imex curation Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. J. Biol. Chem. (0021-9258) 2014 imex curation Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Only protein-protein interactions in vitro In vitro protease assay protease assay predetermined predetermined participant predetermined Yamamoto-Tanaka M., Motoyama A., Miyai M., Matsunaga Y., Matsuda J., Tsuboi R., Hibino T. Affinomics - Interactions curated for the Affinomics consortium. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs and hence results obtained from their use are of dubious biological relevence. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") Cleavage of the peptide-MCA substrate generated fluorescent AMC (7-amino-4-methyl-coumarin), which was subsequently detected. Recombinant saposin A, caspase-14 and derived constructs were determined to be of Human origin by blat of primers given in this publication or in Reference 6. Interactions involving "revC14" or "revC14-Y178" are have not been included as these are artificial constructs. (For revC14 pmid:21183321 states:- "the small subunit was fused to the N-terminal of the large subunit, ending at D146.") toshihiko.hibino1@to.shiseido.co.jp Accepted 2015-FEB-27 by ORCHARD J. Biol. Chem. (0021-9258) 2014 Only protein-protein interactions imex curation p07602-1 Prosaposin PSAP GLBA SAP1 Proactivator polypeptide protein protein human Homo sapiens Human MYALFLLASLLGAALAGPVLGLKECTRGSAVWCQNVKTASDCGAVKHCLQTVWNKPTVKSLPCDICKDVVTAAGDMLKDNATEEEILVYLEKTCDWLPKPNMSASCKEIVDSYLPVILDIIKGEMSRPGEVCSALNLCESLQKHLAELNHQKQLESNKIPELDMTEVVAPFMANIPLLLYPQDGPRSKPQPKDNGDVCQDCIQMVTDIQTAVRTNSTFVQALVEHVKEECDRLGPGMADICKNYISQYSEIAIQMMMHMQPKEICALVGFCDEVKEMPMQTLVPAKVASKNVIPALELVEPIKKHEVPAKSDVYCEVCEFLVKEVTKLIDNNKTEKEILDAFDKMCSKLPKSLSEECQEVVDTYGSSILSILLEEVSPELVCSMLHLCSGTRLPALTVHVTQPKDGGFCEVCKKLVGYLDRNLEKNSTKQEILAALEKGCSFLPDPYQKQCDQFVAEYEPVLIEILVEVMDPSFVCLKIGACPSAHKPLLGTEKCIWGPSYWCQNTETAAQCNAVEHCKRHVWN DIP protein P07602-1 original sequence version: 123 DIP protein P07602 original sequence version: 146 71977F7A8C9E1533 try3_human Trypsin-3 PRSS3 PRSS4 TRY3 TRY4 Trypsin III Brain trypsinogen Mesotrypsinogen Trypsin IV Serine protease 3 Serine protease 4 Mesotrypsin protein protein human Homo sapiens Human MCGPDDRCPARWPGPGRAVKCGKGLAAARPGRVERGGAQRGGAGLELHPLLGGRTWRAARDADGCEALGTVAVPFDDDDKIVGGYTCEENSLPYQVSLNSGSHFCGGSLISEQWVVSAAHCYKTRIQVRLGEHNIKVLEGNEQFINAAKIIRHPKYNRDTLDNDIMLIKLSSPAVINARVSTISLPTTPPAAGTECLISGWGNTLSFGADYPDELKCLDAPVLTQAECKASYPGKITNSMFCVGFLEGGKDSCQRDSGGPVVCNGQLQGVVSWGHGCAWKNRPGVYTKVYNYVDWIKDTIAANS 4C4303C310B7BFFC caspe_human Caspase-14 CASP14 protein protein human Homo sapiens Human MSNPRSLEEEKYDMSGARLALILCVTKAREGSEEDLDALEHMFRQLRFESTMKRDPTAEQFQEELEKFQQAIDSREDPVSCAFVVLMAHGREGFLKGEDGEMVKLENLFEALNNKNCQALRAKPKVYIIQACRGEQRDPGETVGGDEIVMVIKDSPQTIPTYTDALHVYSTVEGYIAYRHDQKGSCFIQTLVDVFTKRKGHILELLTEVTRRMAEAELVQEGKARKTNPEIQSTLRKRLYLQ E539FB7E8DD808A2 p35030-3 Trypsin-3 Mesotrypsin C PRSS3 PRSS4 TRY3 TRY4 Brain trypsinogen Mesotrypsinogen Serine protease 3 Serine protease 4 Trypsin III Trypsin IV protein protein human Homo sapiens Human MNPFLILAFVGAAVAVPFDDDDKIVGGYTCEENSLPYQVSLNSGSHFCGGSLISEQWVVSAAHCYKTRIQVRLGEHNIKVLEGNEQFINAAKIIRHPKYNRDTLDNDIMLIKLSSPAVINARVSTISLPTTPPAAGTECLISGWGNTLSFGADYPDELKCLDAPVLTQAECKASYPGKITNSMFCVGFLEGGKDSCQRDSGGPVVCNGQLQGVVSWGHGCAWKNRPGVYTKVYNYVDWIKDTIAANS 01563656780A6607 boc-gln-ala-arg-methylcoumarin Boc-Gln-Ala-Arg-methylcoumarin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) sap_human Prosaposin PSAP GLBA SAP1 Proactivator polypeptide protein protein human Homo sapiens Human MYALFLLASLLGAALAGPVLGLKECTRGSAVWCQNVKTASDCGAVKHCLQTVWNKPTVKSLPCDICKDVVTAAGDMLKDNATEEEILVYLEKTCDWLPKPNMSASCKEIVDSYLPVILDIIKGEMSRPGEVCSALNLCESLQKHLAELNHQKQLESNKIPELDMTEVVAPFMANIPLLLYPQDGPRSKPQPKDNGDVCQDCIQMVTDIQTAVRTNSTFVQALVEHVKEECDRLGPGMADICKNYISQYSEIAIQMMMHMQPKEICALVGFCDEVKEMPMQTLVPAKVASKNVIPALELVEPIKKHEVPAKSDVYCEVCEFLVKEVTKLIDNNKTEKEILDAFDKMCSKLPKSLSEECQEVVDTYGSSILSILLEEVSPELVCSMLHLCSGTRLPALTVHVTQPKDGGFCEVCKKLVGYLDRNLEKNSTKQEILAALEKGCSFLPDPYQKQCDQFVAEYEPVLIEILVEVMDPSFVCLKIGACPSAHKPLLGTEKCIWGPSYWCQNTETAAQCNAVEHCKRHVWN 71977F7A8C9E1533 p49862-1 Kallikrein-7 Long KLK7 PRSS6 SCCE Serine protease 6 Stratum corneum chymotryptic enzyme protein protein human Homo sapiens Human MARSLLLPLQILLLSLALETAGEEAQGDKIIDGAPCARGSHPWQVALLSGNQLHCGGVLVNERWVLTAAHCKMNEYTVHLGSDTLGDRRAQRIKASKSFRHPGYSTQTHVNDLMLVKLNSQARLSSMVKKVRLPSRCEPPGTTCTVSGWGTTTSPDVTFPSDLMCVDVKLISPQDCTKVYKDLLENSMLCAGIPDSKKNACNGDSGGPLVCRGTLQGLVSWGTFPCGQPNDPGVYTQVCKFTKWINDTMKKHR DIP protein P49862-1 original sequence version: 71 2D68B6B15A76A668 casp14-psap 3 10 unspecified role unspecified role prey prey human-keratinocyte Homo sapiens keratinocyte Keratinocyte 7 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Table 1 -1 This interaction was detected using an extract from differentiated cells - cells 2 days post confluence followed by 1.2 mm calcium treatment and after air exposure. A "growth" phase extract (cells at 80% confluuence) gave no interactors with Pro-caspase-14. casp14-psap-1 3 10 unspecified role unspecified role prey prey human-skin Homo sapiens skin skin 7 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range true ecoli Escherichia coli (strain K12) physical association physical association Table 1 -2 boc_gln_ala_arg_methylcoumarin-prss3 4 8 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 9 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage 2E Enterokinase (of unknown origin) prss3-psap 1 5 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range possible_mature_protein sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain in vitro In vitro 8 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 true 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 true 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 cleavage cleavage reaction cleavage 2F-1 Saposin A-related products were formed by mesotrypsin digestion;other saposin bands were also formed, as were bands from "putative intermediate fragments of higher molecular weight." klk7-prss3 1 11 enzyme enzyme neutral component neutral component 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 5 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range true possible_mature_protein sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain true in vitro In vitro 35 38 cleavage cleavage reaction cleavage 2F-2 KLK7 generated several intermediate products from prosaposin. prss3-psap-1 2 6 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 3C-1 Positive reactions were seen from the lower to upper epidermis, and with a punctuate pattern especially at the stratum granulosum. casp14-prss3 2 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component active_enzyme sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain proximity proximity close-contact colocalization neighbourhood interaction 3C-2 Positive reaction was seen at the stratum granulosum. casp14-prss3-1 2 10 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component active_enzyme sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain true proximity proximity close-contact colocalization neighbourhood interaction 3C-3 Positive reactions were seen at the stratum granulosum and stratum corneum.