Developing approaches to discover protein-protein interactions (PPIs) remains a fundamental challenge. A chemical biology platform is applied here to identify novel PPIs for the AAA+ superfamily oncoprotein reptin. An in silico screen coupled with chemical optimization provided Liddean, a nucleotide-mimetic which modulates reptin's oligomerization status, protein-binding activity and global conformation. Combinatorial peptide phage library screening of Liddean-bound reptin with next generation sequencing identified interaction motifs including a novel reptin docking site on the p53 tumor suppressor protein. Proximity ligation assays demonstrated that endogenous reptin forms a predominantly cytoplasmic complex with its paralog pontin in cancer cells and Liddean promotes a shift of this complex to the nucleus. An emerging view of PPIs in higher eukaryotes is that they occur through a striking diversity of linear peptide motifs. The discovery of a compound that alters reptin's protein interaction landscape potentially leads to novel avenues for therapeutic development.

Centre for Chemical Biology University of Edinburgh EH9 3JG UK Email

Edinburgh Cancer Research Centre Cell Signalling Unit University of Edinburgh EH4 2XR UK Email

RECAMO Masaryk Memorial Cancer Institute 656 53 Brno Czech Republic

School of Chemistry and Biomedical Sciences Research Complex University of St Andrews and EaStCHEM North Haugh St Andrews KY16 9ST UK Email

Zobrazit více v PubMed

Jubb H., Higueruelo A. P., Winter A., Blundell T. L. Trends Pharmacol. Sci. 2012;33:241–248. PubMed

Chen Y.-C., Rajagopala S. V., Stellberger T., Uetz P. Nat. Methods. 2010;7:667–668. PubMed PMC

Lambert J.-P., Ivosev G., Couzens A. L., Larsen B., Taipale M., Lin Z.-Y., Zhong Q., Lindquist S., Vidal M., Aebersold R., Pawson T., Bonner R., Tate S., Gingras A.-C. Nat. Methods. 2013;10:1239–1245. PubMed PMC

Tompa P. Nat. Chem. Biol. 2012;8:597–600. PubMed

Snider J., Thibault G., Houry W. A. Genome Biol. 2008;9:216. PubMed PMC

Stinson B. M., Nager A. R., Glynn S. E., Schmitz K. R., Baker T. A., Sauer R. T. Cell. 2013;153:628–639. PubMed PMC

Nano N., Houry W. Philos. Trans. R. Soc., B. 2013 doi: 10.1098/rstb.2011.0399. PubMed DOI PMC

Shen X., Mizuguchi G., Hamiche A., Wu C. Nature. 2000;406:541–544. PubMed

Jonsson Z. O., Dhar S. K., Narlikar G. J., Auty R., Wagle N., Pellman D., Pratt R. E., Kingston R., Dutta A. J. Biol. Chem. 2001;276:16279–16288. PubMed

Ikura T., Ogryzko V. V., Grigoriev M., Groisman R., Wang J., Horikoshi M., Scully R., Qin J., Nakatani Y. Cell. 2000;102:463–473. PubMed

Doyon Y., Selleck W., Lane W. S., Tan S., Cote J. Mol. Cell. Biol. 2004;24:1884–1896. PubMed PMC

Krogan N. J., Keogh M. C., Datta N., Sawa C., Ryan O. W., Ding H., Haw R. A., Pootoolal J., Tong A., Canadien V., Richards D. P., Wu X., Emili A., Hughes T. R., Buratowski S., Greenblatt J. F. Mol. Cell. 2003;12:1565–1576. PubMed

Cai Y., Jin J., Florens L., Swanson S. K., Kusch T., Li B., Workman J. L., Washburn M. P., Conaway R. C., Conaway J. W. J. Biol. Chem. 2005;280:13665–13670. PubMed

Wood M. A., McMahon S. B., Cole M. D. Mol. Cell. 2000;5:321–330. PubMed

Bauer A., Chauvet S., Huber O., Usseglio F., Rothbacher U., Aragnol D., Kemler R., Pradel J. EMBO J. 2000;19:6121–6130. PubMed PMC

Kim J. H., Kim B., Cai L., Choi H. J., Ohgi K. A., Tran C., Chen C., Chung C. H., Huber O., Rose D. W., Sawyers C. L., Rosenfeld M. G., Baek S. H. Nature. 2005;434:921–926. PubMed

Maslon M. M., Hrstka R., Vojtesek B., Hupp T. R. J. Mol. Biol. 2010;404:418–438. PubMed

Izumi N., Yamashita A., Iwamatsu A., Kurata R., Nakamura H., Saari B., Hirano H., Anderson P., Ohno S. Sci. Signaling. 2010;3:ra27. PubMed

Venteicher A. S., Meng Z., Mason P. J., Veenstra T. D., Artandi S. E. Cell. 2008;132:945–957. PubMed PMC

Gorynia S., Bandeiras T. M., Pinho F. G., McVey C. E., Vonrhein C., Round A., Svergun D. I., Donner P., Matias P. M., Carrondo M. A. J. Struct. Biol. 2011;176:279–291. PubMed

Matias P. M., Gorynia S., Donner P., Carrondo M. A. J. Biol. Chem. 2006;281:38918–38929. PubMed

Petukhov M., Dagkessamanskaja A., Bommer M., Barrett T., Tsaneva I., Yakimov A., Quéval R., Shvetsov A., Khodorkovskiy M., Käs E., Grigoriev M. Structure. 2012;20:1321–1331. PubMed

Cheung K. L. Y., Huen J., Houry W. A., Ortega J. Biochem. Cell Biol. 2010;88:77–88. PubMed PMC

Niewiarowski A., Bradley A. S., Gor J., McKay A. R., Perkins S. J., Tsaneva I. R. Biochem. J. 2010;429:113–125. PubMed

Taylor P., Blackburn E., Sheng Y. G., Harding S., Hsin K.-Y., Kan D., Shave S., Walkinshaw M. D. Br. J. Pharmacol. 2008;153:S55–S67. PubMed PMC

Hernychova L., Man P., Verma C., Nicholson J., Sharma C.-A., Ruckova E., Teo J. Y., Ball K., Vojtesek B., Hupp T. R. Proteomics. 2013;13:2512–2525. PubMed

Stolc V., Samanta M. P., Tongprasit W., Marshall W. F. Proc. Natl. Acad. Sci. U. S. A. 2005;102:3703–3707. PubMed PMC

Zhao L., Yuan S., Cao Y., Kallakuri S., Li Y., Kishimoto N., DiBella L., Sun Z. Proc. Natl. Acad. Sci. U. S. A. 2013;110:12697–12702. PubMed PMC

Clausson C.-M., Allalou A., Weibrecht I., Mahmoudi S., Farnebo M., Landegren U., Wählby C., Söderberg O. Nat. Methods. 2011;8:892–893. PubMed

Queval R., Papin C., Dalvai M., Bystricky K., Humbert O. J. Biol. Chem. 2014;289(49):33999–34012. PubMed PMC

Rosenbaum J., Baek S., Dutta A. Sci. Signaling. 2013;6:1–6. PubMed PMC

Elkaim J., Lamblin M., Laguerre M., Rosenbaum J., Lestienne P., Eloy L., Cresteil T., Felpin F.-X., Dessolin J. Bioorg. Med. Chem. Lett. 2014;24:2512–2516. PubMed

Grigoletto A., Lestienne P., Rosenbaum J. Biochim. Biophys. Acta. 2011;1815:147–157. PubMed

Van Roey K., Uyar B., Weatheritt R. J., Dinkel H., Seiler M., Budd A., Gibson T. J., Davey N. E. Chem. Rev. 2014;114:6733–6778. PubMed

Tompa P., Davey N. E., Gibson T. J., Babu M. M. Mol. Cell. 2014;55:161–169. PubMed

Miller M. L., Jensen L. J., Diella F., Jorgensen C., Tinti M., Li L., Hsiung M., Parker S. A., Bordeaux J., Sicheritz-Ponten T., Olhovsky M., Pasculescu A., Alexander J., Knapp S., Blom N., Bork P., Li S., Cesareni G., Pawson T., Turk B. E., Yaffe M. B., Brunak S., Linding R. Sci. Signaling. 2008;1:ra2. doi: 10.1126/scisignal.1159433. PubMed DOI PMC

Ray-Coquard I., Blay J.-Y., Italiano A., Le Cesne A., Penel N., Zhi J., Heil F., Rueger R., Graves B., Ding M., Geho D., Middleton S. A., Vassilev L. T., Nichols G. L., Bui B. N. Lancet Oncol. 2012;13:1133–1140. PubMed

Basse M. J., Betzi S., Bourgeas R., Bouzidi S., Chetrit B., Hamon V., Morelli X., Roche P. Nucleic Acids Res. 2013;41:D824–D827. PubMed PMC

Dinkel H., Van Roey K., Michael S., Davey N. E., Weatheritt R. J., Born D., Speck T., Krüger D., Grebnev G., Kuban M., Strumillo M., Uyar B., Budd A., Altenberg B., Seiler M., Chemes L. B., Glavina J., Sánchez I. E., Diella F., Gibson T. J. Nucleic Acids Res. 2014;42:D259–D266. PubMed PMC

The Sequence-specific Peptide-binding Activity of the Protein Sulfide Isomerase AGR2 Directs Its Stable Binding to the Oncogenic Receptor EpCAM