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 Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα FEBS Lett. (0014-5793) 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. brigitte.pertschy@uni-graz.at 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-JAN-13 by LUANA 2014 imex curation Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα FEBS Lett. (0014-5793) 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. brigitte.pertschy@uni-graz.at 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-JAN-13 by LUANA 2014 imex curation Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα Ribosomal protein S3 interacts with the NF-κB inhibitor IκBα FEBS Lett. (0014-5793) 2014 imex curation Stanborough T., Niederhauser J., Koch B., Bergler H., Pertschy B. brigitte.pertschy@uni-graz.at 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western brigitte.pertschy@uni-graz.at Accepted 2014-JAN-13 by LUANA 2014 imex curation 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 DIP protein P25963 original sequence version: 160 DIP protein P25963 original sequence version: 157 DIP protein P25963 original sequence version: 145 DIP protein P25963 original sequence version: 170 DIP protein P25963 original sequence version: 122 DIP protein P25963 original sequence version: 128 DIP protein P25963 original sequence version: 78 DIP protein P25963 original sequence version: 121 DIP protein P25963 original sequence version: 124 088B313226786395 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 DIP protein Q04206 original sequence version: 170 DIP protein Q04206 original sequence version: 173 DIP protein Q04206 original sequence version: 99 DIP protein Q04206-1 original sequence version: 99 DIP protein Q04206 original sequence version: 168 DIP protein Q04206 original sequence version: 128 DIP protein Q04206 original sequence version: 131 DIP protein Q04206 original sequence version: 153 DIP protein Q04206 original sequence version: 185 DIP protein Q04206 original sequence version: 181 DIP protein Q04206 original sequence version: 183 8147A6AF9F2445C6 rs3_human 40S ribosomal protein S3 RPS3 Small ribosomal subunit protein uS3 OK/SW-cl.26 protein protein human Homo sapiens Human MAVQISKKRKFVADGIFKAELNEFLTRELAEDGYSGVEVRVTPTRTEIIILATRTQNVLGEKGRRIRELTAVVQKRFGFPEGSVELYAEKVATRGLCAIAQAESLRYKLLGGLAVRRACYGVLRFIMESGAKGCEVVVSGKLRGQRAKSMKFVDGLMIHSGDPVNYYVDTAVRHVLLRQGVLGIKVKIMLPWDPTGKIGPKKPLPDHVSIVEPKDEILPTTPISEQKGGKPEPPAMPQPVPTA DIP protein P23396 original sequence version: 174 6ECBB34A8EE04AAF rps3-nfkbia-5 2 Ikbeta-alpha 4 anti tag western anti tag western blot anti tag western 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 p65 5 anti tag western anti tag western blot anti tag western 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 RPS3 6 anti tag western anti tag western blot anti tag western 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 association association f1b rps3-nfkbia-6 2 Ikbeta-alpha 4 anti tag western anti tag western blot anti tag western 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 RPS3 6 anti tag western anti tag western blot anti tag western 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 physical association physical association f1b nfkbia-rps3 2 RPS3 6 anti tag western anti tag western blot anti tag western 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 Ikbeta-alpha 4 anti tag western anti tag western blot anti tag western 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 physical association physical association f1c nfkbia-rps3-1 1 RPS3 6 anti tag western anti tag western blot anti tag western 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Ikbeta-alpha 4 anti tag western anti tag western blot anti tag western 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 binding region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 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 3 Ikbeta-alpha 4 anti tag western anti tag western blot anti tag western 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 RPS3 6 anti tag western anti tag western blot anti tag western 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 p65 5 western blot western blot Immuno blot Immuno blot anti tag western anti tag western blot anti tag western 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 physical association physical association 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).