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Crystal structure of the blue fluorescent protein with a Leu-Leu-Gly tri-peptide chromophore derived from the purple chromoprotein of Stichodactyla haddoni.
Int J Biol Macromol 2019; 130:675-684IJ

Abstract

Chromoproteins are a good source of engineered biological tools. We previously reported the development of a blue fluorescent protein, termed shBFP, which was derived from a purple chromoprotein shCP found in the sea anemone Stichodacyla haddoni. shBFP contains a Leu63-Leu64-Gly65 tri-peptide chromophore, and shows maximum excitation and emission wavelengths at 401 nm and 458 nm, along with a high quantum yield. How this chromophore endows shBFP with the unique fluorescence property in the absence of a hydroxyphenyl ring remained unclear. Here, we present the crystal structures of shCP and shBFP at 1.9- and 2.05-Å resolution, respectively. Both proteins crystallized as similar tetramers, but they are more likely to function as dimers in solution. The chromophore in shCP shows a trans-conformation and its non-planarity is similar to most other homologues. The shBFP chromophore also contains an imidazolidone moiety in its structure, but there are a smaller number of conjugated double bonds compared to shCP. Consequently, the chromophore may prefer absorbing shorter wavelength lights in the UV region, followed by the emission of blue fluorescence. These observations provide new insights into the molecular basis that correlates chromophore conformation with light absorption and fluorescence emission for the development of improved biomarkers.

Authors+Show Affiliations

Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan; The Asia-Pacific Ocean Research Center, National Sun Yat-sen University, Kaohsiung, Taiwan; Doctoral Degree Program in Marine Biotechnology, National Sun Yat-sen University, Kaohsiung, Taiwan. Electronic address: hychang@mail.nsysu.edu.tw.Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan.Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.Institute of Biomedical Sciences, MacKay Medical College, New Taipei City, Taiwan.Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan.Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.Institute of Biomedical Sciences, MacKay Medical College, New Taipei City, Taiwan. Electronic address: hjtsai@ntu.edu.tw.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30836182

Citation

Chang, Hsin-Yang, et al. "Crystal Structure of the Blue Fluorescent Protein With a Leu-Leu-Gly Tri-peptide Chromophore Derived From the Purple Chromoprotein of Stichodactyla Haddoni." International Journal of Biological Macromolecules, vol. 130, 2019, pp. 675-684.
Chang HY, Ko TP, Chang YC, et al. Crystal structure of the blue fluorescent protein with a Leu-Leu-Gly tri-peptide chromophore derived from the purple chromoprotein of Stichodactyla haddoni. Int J Biol Macromol. 2019;130:675-684.
Chang, H. Y., Ko, T. P., Chang, Y. C., Huang, K. F., Lin, C. Y., Chou, H. Y., ... Tsai, H. J. (2019). Crystal structure of the blue fluorescent protein with a Leu-Leu-Gly tri-peptide chromophore derived from the purple chromoprotein of Stichodactyla haddoni. International Journal of Biological Macromolecules, 130, pp. 675-684. doi:10.1016/j.ijbiomac.2019.02.138.
Chang HY, et al. Crystal Structure of the Blue Fluorescent Protein With a Leu-Leu-Gly Tri-peptide Chromophore Derived From the Purple Chromoprotein of Stichodactyla Haddoni. Int J Biol Macromol. 2019 Jun 1;130:675-684. PubMed PMID: 30836182.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Crystal structure of the blue fluorescent protein with a Leu-Leu-Gly tri-peptide chromophore derived from the purple chromoprotein of Stichodactyla haddoni. AU - Chang,Hsin-Yang, AU - Ko,Tzu-Ping, AU - Chang,Yu-Ching, AU - Huang,Kai-Fa, AU - Lin,Cheng-Yung, AU - Chou,Hong-Yun, AU - Chiang,Cheng-Yi, AU - Tsai,Huai-Jen, Y1 - 2019/03/02/ PY - 2018/12/04/received PY - 2019/02/19/revised PY - 2019/02/23/accepted PY - 2019/3/6/pubmed PY - 2019/8/6/medline PY - 2019/3/6/entrez KW - Biomarker KW - Emission spectra KW - Green fluorescent protein KW - Sea anemone KW - UV absorption KW - X-ray diffraction SP - 675 EP - 684 JF - International journal of biological macromolecules JO - Int. J. Biol. Macromol. VL - 130 N2 - Chromoproteins are a good source of engineered biological tools. We previously reported the development of a blue fluorescent protein, termed shBFP, which was derived from a purple chromoprotein shCP found in the sea anemone Stichodacyla haddoni. shBFP contains a Leu63-Leu64-Gly65 tri-peptide chromophore, and shows maximum excitation and emission wavelengths at 401 nm and 458 nm, along with a high quantum yield. How this chromophore endows shBFP with the unique fluorescence property in the absence of a hydroxyphenyl ring remained unclear. Here, we present the crystal structures of shCP and shBFP at 1.9- and 2.05-Å resolution, respectively. Both proteins crystallized as similar tetramers, but they are more likely to function as dimers in solution. The chromophore in shCP shows a trans-conformation and its non-planarity is similar to most other homologues. The shBFP chromophore also contains an imidazolidone moiety in its structure, but there are a smaller number of conjugated double bonds compared to shCP. Consequently, the chromophore may prefer absorbing shorter wavelength lights in the UV region, followed by the emission of blue fluorescence. These observations provide new insights into the molecular basis that correlates chromophore conformation with light absorption and fluorescence emission for the development of improved biomarkers. SN - 1879-0003 UR - https://www.unboundmedicine.com/medline/citation/30836182/Crystal_structure_of_the_blue_fluorescent_protein_with_a_Leu_Leu_Gly_tri_peptide_chromophore_derived_from_the_purple_chromoprotein_of_Stichodactyla_haddoni_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0141-8130(18)36728-X DB - PRIME DP - Unbound Medicine ER -