Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-27T02:03:32.581Z Has data issue: false hasContentIssue false
  • English
  • Français

Scanning Probe Microscopy of Bacterial Red Light Photoreceptors

Published online by Cambridge University Press:  24 May 2012

Fernando G. Tobias ,
Anna Gawedzka ,
Max S. Goldmeier ,
Alexandra C. Sakols ,
Emina A. Stojković ,
Stefan Tsonchev  and
Kenneth T. Nicholson
Fernando G. Tobias
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Anna Gawedzka
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA Northeastern Illinois University, Department of Biology, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Max S. Goldmeier
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Alexandra C. Sakols
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Emina A. Stojković
Affiliation:
Northeastern Illinois University, Department of Biology, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Stefan Tsonchev
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Kenneth T. Nicholson
Affiliation:
Northeastern Illinois University, Department of Chemistry, 5500 N. St. Louis Ave., Chicago, IL 60625, USA
Get access

Abstract

Bacteriophytochromes (Bphs) are red-light photoreceptors found in photosynthetic and non-photosynthetic bacteria that have been engineered into infrared fluorescent protein markers. Bphs are composed of a photosensory module that is covalently linked to an effector/regulatory module, usually a histidine kinase (HK) domain. Light-induced, global structural changes are proposed to originate within the covalently attached biliverdin chromophore, a linear tetrapyrrole, and propagate through the protein. Bphs undergo reversible photoconversion between two distinct red and far-red light absorbing states, denoted Pr and Pfr respectively. For most Bphs, Pr is the dark-adapted state. The energy dissipated during Pr/Pfr photoconversion is proposed to directly impact the infrared fluorescence quantum yield. At this time, only structures of three different Bphs have been published, all of truncated proteins in their respective dark-adapted states. We have utilized scanning probe microscopy (SPM) to investigate the structure of intact Bphs in the light-adapted state in order to gain new insight into the mechanism of photoconversion and fluorescence. Scanning tunneling microscopy (STM) analysis of a pair of Bphs from photosynthetic bacterium R. palustris, RpBphP2 (P2) and RpBphP3 (P3) in their light-adapted states is presented in these proceedings. The concentration of the depositing protein has a key role in the molecular arrangements observed on the highly-ordered pyrolytic graphite (HOPG) surface. For example, at a high protein concentration, a hexagonal lattice of Bphs is observed by STM on a HOPG surface. Upon dilution, the photoreceptors self-organize into fiber-like structures on the surface. In these fibers, the dimer interface and the individual domains of the Bphs can be assigned and directly compared to a structural model of the intact, full-length proteins. In summary, SPM has potential to be an effective method for gaining new insight into Bph structure and dynamics.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 1465: Symposium SS – Structure/Property Relationships in Biological and Biomimetic Materials at the Micro-, Nano- and Atomic-Length Scales , 2012 , mrss12-1465-ss07-12
Copyright
Copyright © Materials Research Society 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

1. Noack, S., Michael, N., Rosen, R. and Lamparter, T., Biochemistry 46, 4164–4176 (2007).10.1021/bi602419xGoogle Scholar
2. Ponting, C. P. and Aravind, L., Current Biology 7, R674–677 (1997).10.1016/S0960-9822(06)00352-6Google Scholar
3. Rockwell, N. C. and Lagarias, J. C., The Plant Cell 18, 4–14 (2006).10.1105/tpc.105.038513Google Scholar
4. Aravind, L. and Ponting, C. P., Trends in Biochemical Sciences 22, 458–459 (1997).10.1016/S0968-0004(97)01148-1Google Scholar
5. Karniol, B., Wagner, J. R., Walker, J. M. and Vierstra, R. D., Biochemistry Journal 392(Pt 1), 103–116 (2005).10.1042/BJ20050826Google Scholar
6. Noack, S. and Lamparter, T., Methods of Enymology 423, 203–221 (2007).10.1016/S0076-6879(07)23009-5Google Scholar
7. Rockwell, N. C., Su, Y. S. and Lagarias, J. C., Annual Reviews in Plant Biology 57, 837–856 (2006).10.1146/annurev.arplant.56.032604.144208Google Scholar
8. Yang, X., Kuk, J. and Moffat, K., Proceedings of the National Academy of Sciences USA 105, 14715–14720 (2008).10.1073/pnas.0806718105Google Scholar
9. Binnig, G., Rohrer, H., Gerber, C. and Wicbcl, E., Physical Review Letters 49, 57 (1982).10.1103/PhysRevLett.49.57Google Scholar
10. Xia, Chuanjun, Fan, X., Locklin, J., , R. C., Gies, A. A. and Nonidez, W., Journal of the American Chemical Society 126, 8735–8743 (2004).10.1021/ja0484404Google Scholar
11. Satterlee, J. D. and Mazur, U., Journal of Physical Chemistry B 110, 22968–22970 (2006).10.1021/jp065312oGoogle Scholar
12. Wang, Y., Lingenfelder, M., Classen, T., Costantini, G. and Kern, K., Journal of the American Chemical Society 129, 15742–15743 (2007).10.1021/ja075118vGoogle Scholar
13. Ron, I., Sepunaru, L., Itzhakov, S., Belenkova, T., Friedman, N., Pecht, I., Sheves, M. and Cahen, D., Journal of the American Chemical Society 132, 4131–4140 (2010).10.1021/ja907328rGoogle Scholar
14. Giraud, E., Zappa, S., Vuillet, L., Adriano, J.-M., Hannibal, L., Fardoux, J., Berthomieu, C., Bouyer, P., Pignol, D. and Vermeglio, A., The Journal of Biological Chemistry 280, 32389–32397 (2005).10.1074/jbc.M506890200Google Scholar
15. Yang, X., Stojkovic, E. A., Kuk, J. and Moffat, K., Proceedings of the National Academy of Sciences USA 104, 12571–12576 (2007).10.1073/pnas.0701737104Google Scholar
16. Toh, K. C., Stojkovic, E. A., van Stokkum, I. H., Moffat, K. and Kennis, J. T., Proceedings of the National Academy of Sciences USA 107, 9170–9175 (2010).10.1073/pnas.0911535107Google Scholar
17. Filonov, G. S., Piatkevich, K. D., Ting, L.-M., Zhang, J., Kim, K. and Verkhusha, V. V., Nature Biotechnology 29, 757–761 (2011).10.1038/nbt.1918Google Scholar
18. Li, H., Zhang, J., Vierstra, R. and Li, H., Proceedings of the National Academy of Sciences USA 107, 10872–10877 (2010).10.1073/pnas.1001908107Google Scholar
19. Yang, X., Kuk, J. and Moffat, K., Proceedings of the National Academy of Sciences USA 106, 15639–15644 (2009).10.1073/pnas.0902178106Google Scholar
20. The PyMOL Molecular Graphics System, Version 1.5.0.1 Schrödinger, LLC Google Scholar