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/ Soluble interleukin-15 receptor alpha (IL-15R alpha)-sushi as a selective and potent agonist of IL-15 action through IL-15R beta/gamma. Hyperagonist IL-15 x IL-15R alpha fusion proteins. Soluble interleukin-15 receptor alpha (IL-15R alpha)-sushi as a selective and potent agonist of IL-15 action through IL-15R beta/gamma. Hyperagonist IL-15 x IL-15R alpha fusion proteins. J. Biol. Chem. (0021-9258) 2006 imex curation Mortier E., Quemener A., Vusio P., Lorenzen I., Boublik Y., Grotzinger J., Plet A., Jacques Y. BioCreative - Critical Assessment of Information Extraction systems in Biology yjacques@nantes.inserm.fr 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Mortier E., Quemener A., Vusio P., Lorenzen I., Boublik Y., Grotzinger J., Plet A., Jacques Y. BioCreative - Critical Assessment of Information Extraction systems in Biology yjacques@nantes.inserm.fr Accepted 2006-AUG-8 by DAVET. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation i15ra_human Interleukin-15 receptor subunit alpha IL15RA protein protein human Homo sapiens Human MAPRRARGCRTLGLPALLLLLLLRPPATRGITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTAGVTPQPESLSPSGKEPAASSPSSNNTAATTAAIVPGSQLMPSKSPSTGTTEISSHESSHGTPSQTTAKNWELTASASHQPPGVYPQGHSDTTVAISTSTVLLCGLSAVSLLACYLKSRQTPPLASVEMEAMEALPVTWGTSSRDEDLENCSHHL A9CFC885189E96BE il15_human Interleukin-15 IL15 protein protein human Homo sapiens Human MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS 0CE0520C1D8379E2 il15-il15ra 1 2 unspecified role unspecified role bait bait n-terminal sIL-15Ralpha-IL-2 binding region binding-associated region binding region range range certain certain sequence position certain c-terminal human IL-2 - used as a tag for purification fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 3 unspecified role unspecified role prey prey c-terminal recombinant humanIL-15 from Peprotech Inc. sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association The recombinant IL-15 from Peprotech Inc. corresponds to residues 48-162 of the Uniprot entry except at position 48 there is met substituted for ala. 1 A As shown in Fig. 1A, a sIL-15R IL-2 fusion protein produced in Chinese hamster ovary cells and consisting of the entire IL-15R extracellular domain linked to a molecule of human IL-2 (used as a tag for purification) bound IL-15 with high affinity (kon 3.7 105 M 1 s 1;koff 1.4 10 5 s 1; Kd 38 pM). kon =3.7 E5 per M per S; koff =1.4 E-5 per s; Kd=3.8 E-11 M These results indicate that the sushi domain is responsible for most of the binding affinity of IL-15 but that it does not fully reconstitute the high affinity binding displayed by the full-length extracellular domain. The IL-15Ralpha proteins are annotated to begin at residue 31 as the Uniprot annotation indicates residues 1-30 constitute a signal peptide, which should be removed when expressed in CHO cells. il15-il15ra-1 1 3 unspecified role unspecified role prey prey c-terminal recombinant humanIL-15 from Peprotech Inc. sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 2 unspecified role unspecified role bait bait c-terminal human IL-2 - used as a tag for purification fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range n-terminal sIL-15Ralpha-sushi-IL-2 binding region binding-associated region binding region range range range range crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. physical association physical association The recombinant IL-15 from Peprotech Inc. corresponds to residues 48-162 of the Uniprot entry except at position 48 there is met substituted for ala. 1 B A similar construction linking the sushi domain of IL-15R to human IL-2 also bound IL-15 (Fig. 1B) but with a 10-fold lower affinity, mainly due to a more rapid off rate (kon 3.1 105 M1 s 1; koff 1.3 10 4 s 1; Kd 420 pM). kon=3.1 E5 per M per s; koff =1.3 E-4 per s; Kd =4.2 E-10 M These results indicate that the sushi domain is responsible for most of the binding affinity of IL-15 but that it does not fully reconstitute the high affinity binding displayed by the full-length extracellular domain. The IL-15Ralpha proteins are annotated to begin at residue 31 as the Uniprot annotation indicates residues 1-30 constitute a signal peptide, which should be removed when expressed in CHO cells. il15-il15ra-2 1 3 unspecified role unspecified role prey prey c-terminal recombinant humanIL-15 from Peprotech Inc. sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 2 unspecified role unspecified role bait bait n-terminal sIL-15Ralpha-sushi binding region binding-associated region binding region range range range range ecoli Escherichia coli (strain K12) physical association physical association The recombinant IL-15 from Peprotech Inc. corresponds to residues 48-162 of the Uniprot entry except at position 48 there is met substituted for ala. 1 C A soluble sushi domain was also produced in E. coli. This sIL-15R-sushi also bound IL-15 with a lower affinity (kon 2.5 105 M 1 s 1; koff 3.8 10 4 s 1; Kd 1.5 nM) (Fig. 1C). kon =2.5 E5 per M per s ; koff=3.8 E-4 per s; Kd= 1.5 E-9 M It is not clear if the signal peptide (Uniprot annotation indicates it is residues 1-30 of the sequence) is removed from the sIL-15Ralpha-sushi expressed in E. coli (referred to in the annotation topic kinetics). Nor is it clear if this protein is fused to human Il-2, but it seems not as IL-2 is not included in the name of the construct. These results indicate that the sushi domain is responsible for most of the binding affinity of IL-15 but that it does not fully reconstitute the high affinity binding displayed by the full-length extracellular domain. The IL-15Ralpha proteins are annotated to begin at residue 31 as the Uniprot annotation indicates residues 1-30 constitute a signal peptide, which should be removed when expressed in CHO cells.