Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Photo-cross-linking interacting proteins with a genetically encoded benzophenone

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Structure of pBpa and a schematic of the pDULE plasmid.
Figure 2: The steps to express a protein with a site-specifically incorporated pBpa in E. coli.
Figure 3: Analysis of pBpa-containing protein production.
Figure 4: Irradiation time courses of HDH-225pBpa and HDH-225Tyr.

References

  1. Fields, S. & Song, O. A novel genetic system to detect protein-protein interactions. Nature 340, 245–246 (1989).

    CAS  Article  Google Scholar 

  2. Uetz, P. Two-hybrid arrays. Curr. Opin. Chem. Biol. 6, 57–62 (2002).

    CAS  Article  Google Scholar 

  3. Gavin, A.C. et al. Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature 415, 141–147 (2002).

    CAS  Article  Google Scholar 

  4. Ho, Y. et al. Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry. Nature 415, 180–183 (2002).

    CAS  Article  Google Scholar 

  5. Kauer, J.C., Erickson-Viitanen, S., Wolfe, H.R. & DeGrado, W.F. p-benzoyl-L-phenylalanine, a new photoreactive amino acid. J. Biol. Chem. 261, 10695–10700 (1986).

    CAS  PubMed  Google Scholar 

  6. Dorman, G. & Prestwich, G.D. Benzophenone photophores in biochemistry. Biochemistry 33, 5661–5673 (1994).

    CAS  Article  Google Scholar 

  7. Chin, J.W., Martin, A.B., King, D.S., Wang, L. & Schultz, P.G. Addition of a photocrosslinking amino acid to the genetic code of Escherichia coli. Proc. Natl. Acad. Sci. 99, 11020–11024 (2002).

    CAS  Article  Google Scholar 

  8. Chin, J.W. & Schultz, P.G. In vivo photocrosslinking with unnatural amino acid mutagenesis. Chembiochem. 11, 1135–1137 (2002).

    Article  Google Scholar 

  9. Schlieker, C. et al. Substrate recognition by the AAA+ chaperone ClpB. Nat. Struct. Mol. Biol. 11, 607–615 (2004).

    CAS  Article  Google Scholar 

  10. Pryor, K.D. & Leiting, B. High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding-protein double-affinity fusion system. Protein Expr. Purif. 10, 309–319 (1997).

    CAS  Article  Google Scholar 

  11. Miller, J.H. & Albertini, A.M. Effects of surrounding sequence on the suppression of nonsense codons. J. Mol. Biol. 164, 59–71 (1983).

    CAS  Article  Google Scholar 

  12. Chin, J.W. et al. An expanded eukaryotic genetic code. Science 301, 964–967 (2003).

    CAS  Article  Google Scholar 

  13. Hino N. et al. Protein photo-cross-linking in mammalian cells by site-specific incorporation of a photoreactive amino acid. Nat. Methods 2, 201–206 (2005).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

J.W.C. and R.A.M. thank Peter Schultz for his support in the development of some of the vectors described in this protocol.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Ryan A Mehl or Jason W Chin.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Farrell, I., Toroney, R., Hazen, J. et al. Photo-cross-linking interacting proteins with a genetically encoded benzophenone. Nat Methods 2, 377–384 (2005). https://doi.org/10.1038/nmeth0505-377

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth0505-377

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing