A Bifunctional Amino Acid Enables Both Covalent Chemical Capture and Isolation of in Vivo Protein–Protein Interactions
dc.contributor.author | Joiner, Cassandra M. | |
dc.contributor.author | Breen, Meghan E. | |
dc.contributor.author | Clayton, James | |
dc.contributor.author | Mapp, Anna K. | |
dc.date.accessioned | 2017-02-02T22:00:12Z | |
dc.date.available | 2018-03-01T16:43:50Z | en |
dc.date.issued | 2017-01-17 | |
dc.identifier.citation | Joiner, Cassandra M.; Breen, Meghan E.; Clayton, James; Mapp, Anna K. (2017). "A Bifunctional Amino Acid Enables Both Covalent Chemical Capture and Isolation of in Vivo Protein–Protein Interactions." ChemBioChem 18(2): 181-184. | |
dc.identifier.issn | 1439-4227 | |
dc.identifier.issn | 1439-7633 | |
dc.identifier.uri | https://hdl.handle.net/2027.42/135955 | |
dc.description.abstract | In vivo covalent chemical capture by using photoactivatable unnatural amino acids (UAAs) is a powerful tool for the identification of transient protein–protein interactions (PPIs) in their native environment. However, the isolation and characterization of the crosslinked complexes can be challenging. Here, we report the first in vivo incorporation of the bifunctional UAA BPKyne for the capture and direct labeling of crosslinked protein complexes through post‐crosslinking functionalization of a bioorthogonal alkyne handle. Using the prototypical yeast transcriptional activator Gal4, we demonstrate that BPKyne is incorporated at the same level as the commonly used photoactivatable UAA pBpa and effectively captures the Gal4–Gal80 transcriptional complex. Post‐crosslinking, the Gal4–Gal80 adduct was directly labeled by treatment of the alkyne handle with a biotin‐azide probe; this enabled facile isolation and visualization of the crosslinked adduct from whole‐cell lysate. This bifunctional amino acid extends the utility of the benzophenone crosslinker and expands our toolbox of chemical probes for mapping PPIs in their native cellular environment.Using the bifunctional unnatural amino acid, BPKyne, we have developed a strategy to capture and directly label transient protein–protein interactions (PPIs) in their native environment. Click chemical functionalization post‐crosslinking with a biotin–azide probe enabled the isolation of transcriptional protein complexes from yeast cells. This amino acid will expand the toolbox for the discovery of new PPIs in live cells. | |
dc.publisher | Academic Press | |
dc.publisher | Wiley Periodicals, Inc. | |
dc.subject.other | protein–protein interactions | |
dc.subject.other | click chemistry | |
dc.subject.other | bioorthogonal labeling | |
dc.subject.other | unnatural amino acids | |
dc.subject.other | photo-crosslinking | |
dc.title | A Bifunctional Amino Acid Enables Both Covalent Chemical Capture and Isolation of in Vivo Protein–Protein Interactions | |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | |
dc.subject.hlbsecondlevel | Biological Chemistry | |
dc.subject.hlbtoplevel | Health Sciences | |
dc.subject.hlbtoplevel | Science | |
dc.description.peerreviewed | Peer Reviewed | |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/135955/1/cbic201600578.pdf | |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/135955/2/cbic201600578_am.pdf | |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/135955/3/cbic201600578-sup-0001-misc_information.pdf | |
dc.identifier.doi | 10.1002/cbic.201600578 | |
dc.identifier.source | ChemBioChem | |
dc.identifier.citedreference | C. E. Fritze, T. R. Anderson in Methods in Enzymology, Vol. 327: Applications of Chimeric Genes and Hybrid Proteins–Part B: Cell Biology and Physiology (Eds.: J. Thorner, S. D. Emr, J. N. Abelson ), Academic Press, San Diego, 2000, pp. 3 – 16. | |
dc.identifier.citedreference | J. R. Perkins, I. Diboun, B. H. Dessailly, J. G. Lees, C. Orengo, Structure 2010, 18, 1233 – 1243; | |
dc.identifier.citedreference | T. Berggård, S. Linse, P. James, Proteomics 2007, 7, 2833 – 2842. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | J. W. Chin, P. G. Schultz, ChemBioChem 2002, 3, 1135 – 1137; | |
dc.identifier.citedreference | J. W. Chin, T. A. Cropp, J. C. Anderson, M. Mukherji, Z. Zhang, P. G. Schultz, Science 2003, 301, 964 – 967; | |
dc.identifier.citedreference | W. Liu, L. Alfonta, A. V. Mack, P. G. Schultz, Angew. Chem. Int. Ed. 2007, 46, 6073 – 6075; Angew. Chem. 2007, 119, 6185 – 6187; | |
dc.identifier.citedreference | Q. Wang, L. Wang, J. Am. Chem. Soc. 2008, 130, 6066 – 6067; | |
dc.identifier.citedreference | T. C. Lee, M. Kang, C. H. Kim, P. G. Schultz, E. Chapman, A. A. Deniz, ChemBioChem 2016, 17, 981 – 984; | |
dc.identifier.citedreference | A. Yamaguchi, T. Matsuda, K. Ohtake, T. Yanagisawa, S. Yokoyama, Y. Fujiwara, T. Watanabe, T. Hohsaka, K. Sakamoto, Bioconjugate Chem. 2016, 27, 198 – 206; | |
dc.identifier.citedreference | N. Hino, Y. Okazaki, T. Kobayashi, A. Hayashi, K. Sakamoto, S. Yokoyama, Nat. Methods 2005, 2, 201 – 206. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | C. Y. Majmudar, L. W. Lee, J. K. Lancia, A. Nwokoye, Q. Wang, A. M. Wangs, L. Wang, A. K. Mapp, J. Am. Chem. Soc. 2009, 131, 14240 – 14242; | |
dc.identifier.citedreference | M. Krishnamurthy, A. Dugan, A. Nwokoye, Y.-H. Fung, J. K. Lancia, C. Y. Majmudar, A. K. Mapp, ACS Chem. Biol. 2011, 6, 1321 – 1326; | |
dc.identifier.citedreference | A. Dugan, R. Pricer, M. Katz, A. K. Mapp, Protein Sci. 2016, 25, 1371 – 1377. | |
dc.identifier.citedreference | S. I. Presolski, V. P. Hong, M. G. Finn in Current Protocols in Chemical Biology, Wiley, Hoboken, 2009. | |
dc.identifier.citedreference | Y. Chen, Y. Wu, P. Henklein, X. Li, K. P. Hofmann, K. Nakanishi, O. P. Ernst, Chem. Eur. J. 2010, 16, 7389 – 7394. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | J. B. Thoden, L. A. Ryan, R. J. Reece, H. M. Holden, J. Biol. Chem. 2008, 283, 30266 – 30272; | |
dc.identifier.citedreference | A. Z. Ansari, R. J. Reece, M. Ptashne, Proc. Natl. Acad. Sci. USa 1998, 95, 13543 – 13548. | |
dc.identifier.citedreference | A. Ahlburg, A. T. Lindhardt, R. H. Taaning, A. E. Modvig, T. Skrydstrup, J. Org. Chem. 2013, 78, 10310 – 10318. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | G. Dorman, G. D. Prestwich, Biochemistry 1994, 33, 5661 – 5673. | |
dc.identifier.citedreference | J. K. Lancia, A. Nwokoye, A. Dugan, C. Joiner, R. Pricer, A. K. Mapp, Biopolymers 2014, 101, 391 – 397. | |
dc.identifier.citedreference | A. L. Stokes, S. J. Miyake-Stoner, J. C. Peeler, D. P. Nguyen, R. P. Hammer, R. A. Mehl, Mol. BioSyst. 2009, 5, 1032; | |
dc.identifier.citedreference | Y.-S. Wang, X. Fang, A. L. Wallace, B. Wu, W. R. Liu, J. Am. Chem. Soc. 2012, 134, 2950 – 2953; | |
dc.identifier.citedreference | D. D. Young, S. Jockush, N. J. Turro, P. G. Schultz, Bioorg. Med. Chem. Lett. 2011, 21, 7502 – 7504. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | A. L. Hopkins, C. R. Groom, Nat. Rev. Drug Discovery 2002, 1, 727 – 730; | |
dc.identifier.citedreference | S. Surade, T. L. Blundell, Chem. Biol. 2012, 19, 42 – 50. | |
dc.identifier.citedreference | ||
dc.identifier.citedreference | A. D. Thompson, A. Dugan, J. E. Gestwicki, A. K. Mapp, ACS Chem. Biol. 2012, 7, 1311 – 1320; | |
dc.identifier.citedreference | A. K. Mapp, A. Z. Ansari, ACS Chem. Biol. 2007, 2, 62 – 75; | |
dc.identifier.citedreference | J.-F. Rual, K. Venkatesan, T. Hao, T. Hirozane-Kishikawa, A. Dricot, N. Li, G. F. Berriz, F. D. Gibbons, M. Dreze, N. Ayivi-Guedehoussou, et al., Nature 2005, 437, 1173 – 1178; | |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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