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 anonymous-2013v-1 Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western brigitte.pertschy@uni-graz.at 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. FEBS Lett. (0014-5793) Only protein-protein interactions anonymous-2013q-2 Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα ecoli Escherichia coli (strain K12) anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western brigitte.pertschy@uni-graz.at 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. FEBS Lett. (0014-5793) Only protein-protein interactions anonymous-2013l-4 Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western brigitte.pertschy@uni-graz.at 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. FEBS Lett. (0014-5793) Only protein-protein interactions ikba_human NF-kappa-B inhibitor alpha NFKBIA IKBA MAD3 NFKBI I-kappa-B-alpha Major histocompatibility complex enhancer-binding protein MAD3 protein protein human Homo sapiens Human MFQAAERPQEWAMEGPRDGLKKERLLDDRHDSGLDSMKDEEYEQMVKELQEIRLEPQEVPRGSEPWKQQLTEDGDSFLHLAIIHEEKALTMEVIRQVKGDLAFLNFQNNLQQTPLHLAVITNQPEIAEALLGAGCDPELRDFRGNTPLHLACEQGCLASVGVLTQSCTTPHLHSILKATNYNGHTCLHLASIHGYLGIVELLVSLGADVNAQEPCNGRTALHLAVDLQNPDLVSLLLKCGADVNRVTYQGYSPYQLTWGRPSTRIQQQLGQLTLENLQMLPESEDEESYDTESEFTEFTEDELPYDDCVFGGQRLTL tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS rs3_human 40S ribosomal protein S3 RPS3 Small ribosomal subunit protein uS3 OK/SW-cl.26 protein protein human Homo sapiens Human MAVQISKKRKFVADGIFKAELNEFLTRELAEDGYSGVEVRVTPTRTEIIILATRTQNVLGEKGRRIRELTAVVQKRFGFPEGSVELYAEKVATRGLCAIAQAESLRYKLLGGLAVRRACYGVLRFIMESGAKGCEVVVSGKLRGQRAKSMKFVDGLMIHSGDPVNYYVDTAVRHVLLRQGVLGIKVKIMLPWDPTGKIGPKKPLPDHVSIVEPKDEILPTTPISEQKGGKPEPPAMPQPVPTA rps3-nfkbia-5 1477165 n/a Ikbeta-alpha 1477168 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a p65 1477169 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 n/a RPS3 1477170 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 association association false false false f1b rps3-nfkbia-6 1477165 n/a Ikbeta-alpha 1477168 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a RPS3 1477170 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 physical association physical association false false false f1b nfkbia-rps3 1477165 n/a RPS3 1477170 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 Ikbeta-alpha 1477168 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f1c nfkbia-rps3-1 1477166 n/a RPS3 1477170 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a Ikbeta-alpha 1477168 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 binding region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false f2 f3c Strikingly, pull-down of IκBα resulted in co-purification of significant amounts of RPS3 (Fig. 2). However, very little RPS3 co-purified with p65(1-311). These data clearly demonstrate a strong direct interaction between RPS3 and IκBα. In contrast, the weak binding of RPS3 to p65 suggests that these two proteins engage in a rather transient interaction. rela-nfkbia-10 1477167 n/a Ikbeta-alpha 1477168 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component partially purified partially purified 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 RPS3 1477170 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component partially purified partially purified 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 n/a p65 1477169 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false f4b The Flag-IκBα-6xHis-RPS3 complex was purified via His-tag purification of 6xHis-RPS3 followed by binding of the His-eluate to ANTIFLAG ® M2 affinity gel (Sigma). The immobilized Flag-IκBα-6xHis-RPS3 complex as well as anti-FLAG agarose bound Flag-IκBα and anti-FLAG agarose bound Flag- RPS3 were incubated with an approximately three-fold molar excess of p65 or with buffer containing TEV-protease (control). Following washing of the beads in washing buffer, bound proteins were eluted by competition with FLAG Peptide (Sigma).