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Ultrafast excited-state dynamics in the green fluorescent protein variant S65T/H148D. 1. Mutagenesis and structural studies.
Biochemistry. 2007 Oct 30; 46(43):12005-13.B

Abstract

Wild type green fluorescent protein (wt-GFP) and the variant S65T/H148D each exhibit two absorption bands, A and B, which are associated with the protonated and deprotonated chromophores, respectively. Excitation of either band leads to green emission. In wt-GFP, excitation of band A (approximately 395 nm) leads to green emission with a rise time of 10-15 ps, due to excited-state proton transfer (ESPT) from the chromophore hydroxyl group to an acceptor. This process produces an anionic excited-state intermediate I* that subsequently emits a green photon. In the variant S65T/H148D, the A band absorbance maximum is red-shifted to approximately 415 nm, and as detailed in the accompanying papers, when the A band is excited, green fluorescence appears with a rise time shorter than the instrument time resolution (approximately 170 fs). On the basis of the steady-state spectroscopy and high-resolution crystal structures of several variants described herein, it is proposed that in S65T/H148D, the red shift of absorption band A and the ultrafast appearance of green fluorescence upon excitation of band A are due to a very short (<or=2.4 A), and possibly low-barrier, hydrogen bond between the chromophore hydroxyl and introduced Asp148.

Authors+Show Affiliations

Institute of Molecular Biology and Department of Physics, University of Oregon, Eugene, Oregon 97403-1229, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

17918959

Citation

Shu, Xiaokun, et al. "Ultrafast Excited-state Dynamics in the Green Fluorescent Protein Variant S65T/H148D. 1. Mutagenesis and Structural Studies." Biochemistry, vol. 46, no. 43, 2007, pp. 12005-13.
Shu X, Kallio K, Shi X, et al. Ultrafast excited-state dynamics in the green fluorescent protein variant S65T/H148D. 1. Mutagenesis and structural studies. Biochemistry. 2007;46(43):12005-13.
Shu, X., Kallio, K., Shi, X., Abbyad, P., Kanchanawong, P., Childs, W., Boxer, S. G., & Remington, S. J. (2007). Ultrafast excited-state dynamics in the green fluorescent protein variant S65T/H148D. 1. Mutagenesis and structural studies. Biochemistry, 46(43), 12005-13.
Shu X, et al. Ultrafast Excited-state Dynamics in the Green Fluorescent Protein Variant S65T/H148D. 1. Mutagenesis and Structural Studies. Biochemistry. 2007 Oct 30;46(43):12005-13. PubMed PMID: 17918959.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ultrafast excited-state dynamics in the green fluorescent protein variant S65T/H148D. 1. Mutagenesis and structural studies. AU - Shu,Xiaokun, AU - Kallio,Karen, AU - Shi,Xinghua, AU - Abbyad,Paul, AU - Kanchanawong,Pakorn, AU - Childs,William, AU - Boxer,Steven G, AU - Remington,S James, Y1 - 2007/10/06/ PY - 2007/10/9/pubmed PY - 2008/1/11/medline PY - 2007/10/9/entrez SP - 12005 EP - 13 JF - Biochemistry JO - Biochemistry VL - 46 IS - 43 N2 - Wild type green fluorescent protein (wt-GFP) and the variant S65T/H148D each exhibit two absorption bands, A and B, which are associated with the protonated and deprotonated chromophores, respectively. Excitation of either band leads to green emission. In wt-GFP, excitation of band A (approximately 395 nm) leads to green emission with a rise time of 10-15 ps, due to excited-state proton transfer (ESPT) from the chromophore hydroxyl group to an acceptor. This process produces an anionic excited-state intermediate I* that subsequently emits a green photon. In the variant S65T/H148D, the A band absorbance maximum is red-shifted to approximately 415 nm, and as detailed in the accompanying papers, when the A band is excited, green fluorescence appears with a rise time shorter than the instrument time resolution (approximately 170 fs). On the basis of the steady-state spectroscopy and high-resolution crystal structures of several variants described herein, it is proposed that in S65T/H148D, the red shift of absorption band A and the ultrafast appearance of green fluorescence upon excitation of band A are due to a very short (<or=2.4 A), and possibly low-barrier, hydrogen bond between the chromophore hydroxyl and introduced Asp148. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/17918959/Ultrafast_excited_state_dynamics_in_the_green_fluorescent_protein_variant_S65T/H148D__1__Mutagenesis_and_structural_studies_ L2 - https://dx.doi.org/10.1021/bi7009037 DB - PRIME DP - Unbound Medicine ER -