Skip to main content
Log in

Structural Dynamics of a Single Photoreceptor Protein Molecule Monitored With Surface-Enhanced Raman Scattering Substrates

  • Published:
MRS Online Proceedings Library Aims and scope

Abstract

Photoactive yellow protein (PYP) is a small cytosolic photoreceptor that actuates the negative phototactic response in its host organism Halorhodospira halophila. It has an optical absorption maximum at 446 nm (blue light). We report an initial study of the photocycle of PYP at the single molecule level using “high enhancement factor” surface-enhanced Raman scattering (SERS)-active nanostructures with 514 nm laser excitation. The SERS-active “nanometal-on-semiconductor” structures are prepared employing a redox technique on thin germanium films, coated on glass slides. Single molecule spectra are observed in terms of sudden appearance of discernable Raman peaks with spectral fluctuations. The single molecule spectra capture protonation, photo-isomerization, and H-bond breaking - the steps that are instrumental in the photocycle of PYP. This is indicative of single PYP molecules diffusing to high-enhancement-factor SERS sites, and undergoing photo-cycle under 514 nm excitation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. T.E. Meyer , Biochim. Biophys. Acta 806, 175 (1985).

    Google Scholar 

  2. W.W. Sprenger , W.D. Hoff , J.P. Armitage , K.J. Hellingwerf , J. Bacteriol. 175, 3096 (1993).

    Article  CAS  Google Scholar 

  3. W.D. Hoff , P. Dux , K. Hard , B. Devreese , I.M. Nugteren-Roodzant , W. Crielaard , R. Boelens , R. Kaptein , J. Van Beeumen , K.J. Hellingwerf , Biochemistry 33, 13959 (1994).

    Article  CAS  Google Scholar 

  4. M. Baca , G.E.O. Borgstahl , M. Boissinot , P.M. Burke , D.R. Williams , K.A. Slater , E.D. Getzoff , Biochemistry 22, 14369 (1994).

    Article  Google Scholar 

  5. T.E. Meyer , E. Yakali , M.A. Cusanovich , G. Tollin , Biochemistry 26, 418 (1987).

    Article  CAS  Google Scholar 

  6. W.D. Hoff , I.H.M. Van Stokkum , H.J. Van Ramesdonk , M.E. Van Brederode , A.M. Brouwer , J.C. Fitch , T.E. Meyer , R. Van Grondelle , K.J. Hellingwerf , Biophys. J. 67, 1691 (1994b).

    Article  CAS  Google Scholar 

  7. M. Kim , R.A. Mathies , W.D. Hoff , K.J. Hellingwerf , Biochemistry 34, 12669 (1995).

    Article  CAS  Google Scholar 

  8. A. Xie , W.D. Hoff , A.R. Kroon , K.J. Hellingwerf , Biochemistry 35, 14671 (1996).

    Article  CAS  Google Scholar 

  9. G.E.O. Borgstahl , D.R. Williams , E.D. Getzoff , Biochemistry 34, 6278 (1995).

    Article  CAS  Google Scholar 

  10. M. Unno , M. Kumauchi , J. Sasaki , F. Tokunaga , S. Yamauchi , Biochemistry 41, 5668 (2002).

    Article  CAS  Google Scholar 

  11. A. Xie , L. Kelemen , J. Hendriks , B.J. White , K.J. Hellingwerf , W.D. Hoff , Biochemistry 40, 1510 (2001).

    Article  CAS  Google Scholar 

  12. B.C. Lee , A. Pandit , P.A. Croonquist , W.D. Hoff , Proc. Natl. Acad. Sci. USA 98, 9062 (2001).

    Article  CAS  Google Scholar 

  13. S. Nie and S.R. Emory , Science 275, 1102 (1997).

    Article  CAS  Google Scholar 

  14. K.J. Hellingwerf , J. Hendriks , T. Gensch , Journal of Physical Chemistry A 107, 1082 (2003).

    Article  CAS  Google Scholar 

  15. M.A. Cusanovich and T.E Meyer . Biochemistry 42, 4759 (2003).

    Article  CAS  Google Scholar 

  16. A.K. Kalkan and S.J. Fonash , J. Phys. Chem. B 109, 20779 (2005).

    Article  Google Scholar 

  17. M. Unno , M. Kumauchi , J. Sasaki , F. Tokunaga , S. Yamauchi , J. Phys. Chem. B 107, 2837 (2003).

    Article  Google Scholar 

  18. M. Unno , M. Kumauchi , F. Tokunaga , S. Yamauchi , J. Phys. Chem. B 111, 2719 (2007).

    Article  Google Scholar 

  19. D. Pan , A. Phillip , W.D. Hoff , R.A. Mathies , Biophysical Journal 86, 2374 (2004).

    Article  CAS  Google Scholar 

  20. A.A. Adesokan , D. Pan , E. Fredj , R.A. Mathies , R.B. Gerber , J. Am. Chem. Soc. 129, 4584 (2007).

    Article  CAS  Google Scholar 

  21. J. M. Zhao , H. Lee , R.A. Nome , S. Majid , N.F. Scherer , W.D. Hoff , Proc. Natl. Acad. Sci. USA 103, 11561 (2006).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Singhal, K., Bhatt, K., Kang, Z. et al. Structural Dynamics of a Single Photoreceptor Protein Molecule Monitored With Surface-Enhanced Raman Scattering Substrates. MRS Online Proceedings Library 1077, 8 (2008). https://doi.org/10.1557/PROC-1077-L10-04

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1557/PROC-1077-L10-04

Navigation