This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We propose continued structural studies on the basis of fluorescence photoswitching in mTFP0.7, a monomeric cyan fluorescent protein distantly related to the green fluorescent protein (GFP). mTFP0.7 is initially brightly fluorescent with a peak emission wavelength of 488 nm, however, rapid photoswitching to a nonfluorescent form occurs when the protein is illuminated with blue light (~ 450 nm)[1]. Spontaneous fluorescence recovery to nearly 100% is observed on a time scale of about 8 minutes at room temperature. Photoswitched mTFP0.7 can be very rapidly reactivated by exposure to 360-400 nm light. Structures of mTFP0.7 have been solved in the fluorescent and cryo-trapped nonfluorescent states[2]. These suggest a mechanism for the change in emission characteristics. In the fluorescent state, the chromophore is cis-coplanar (as found in the vast majority of brightly fluorescent proteins), however, in the dark nonfluorescent state, the chromophore is trans, highly twisted, protonated and partially disordered. We concluded that the latter three characteristics could each be argued to lower emission efficiency and taken together provide a compelling explanation for the complete loss of fluorescence. These characteristics provide a useful model for photoswitching, applicable to all photoswitchable fluorescent proteins described to date. Furthermore, a dramatic rearrangement of internal side chains suggests a ?lock and key? mechanism to explain the long life of the nonfluorescent state. The proposed time-resolved crystallographic studies on mTFP0.7 seek to reveal the basis for the photoswitching phenomenon.
| Weingarten, Adam S; Dannenhoffer, Adam J; Kazantsev, Roman V et al. (2018) Chromophore Dipole Directs Morphology and Photocatalytic Hydrogen Generation. J Am Chem Soc 140:4965-4968 |
| Yang, Cheolhee; Choi, Minseo; Kim, Jong Goo et al. (2018) Protein Structural Dynamics of Wild-Type and Mutant Homodimeric Hemoglobin Studied by Time-Resolved X-Ray Solution Scattering. Int J Mol Sci 19: |
| Kazantsev, Roman V; Dannenhoffer, Adam J; Weingarten, Adam S et al. (2017) Crystal-Phase Transitions and Photocatalysis in Supramolecular Scaffolds. J Am Chem Soc 139:6120-6127 |
| Fournier, Bertrand; Sokolow, Jesse; Coppens, Philip (2016) Analysis of multicrystal pump-probe data sets. II. Scaling of ratio data sets. Acta Crystallogr A Found Adv 72:250-60 |
| Cho, Hyun Sun; Schotte, Friedrich; Dashdorj, Naranbaatar et al. (2016) Picosecond Photobiology: Watching a Signaling Protein Function in Real Time via Time-Resolved Small- and Wide-Angle X-ray Scattering. J Am Chem Soc 138:8815-23 |
| Pande, Kanupriya; Hutchison, Christopher D M; Groenhof, Gerrit et al. (2016) Femtosecond structural dynamics drives the trans/cis isomerization in photoactive yellow protein. Science 352:725-9 |
| Weingarten, Adam S; Kazantsev, Roman V; Palmer, Liam C et al. (2015) Supramolecular Packing Controls H? Photocatalysis in Chromophore Amphiphile Hydrogels. J Am Chem Soc 137:15241-6 |
| Pfoh, Roland; Pai, Emil F; Saridakis, Vivian (2015) Nicotinamide mononucleotide adenylyltransferase displays alternate binding modes for nicotinamide nucleotides. Acta Crystallogr D Biol Crystallogr 71:2032-9 |
| Mariette, Céline; Guérin, Laurent; Rabiller, Philippe et al. (2015) The creation of modulated monoclinic aperiodic composites in n-alkane/urea compounds. Z Kristallogr Cryst Mater 230:5-11 |
| Yang, Xiaojing; Stojkovi?, Emina A; Ozarowski, Wesley B et al. (2015) Light Signaling Mechanism of Two Tandem Bacteriophytochromes. Structure 23:1179-89 |
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