Spectral and structural analysis of large Stokes shift fluorescent protein dKeima570

Yongbin Xu; Kwang Yeon Hwang; Ki Hyun Nam

doi:10.1007/s12275-018-8319-5

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Spectral and structural analysis of large Stokes shift fluorescent protein dKeima570: Yongbin Xu ^1,2, Kwang Yeon Hwang ³, Ki Hyun Nam ^3,4; Journal of Microbiology 2018;56(11):822-827
DOI: https://doi.org/10.1007/s12275-018-8319-5
Published online: October 24, 2018

¹Department of Bioengineering, College of Life Science, Dalian Minzu University, Dalian 116600, P. R. China, ²Key Laboratory of Biotechnology and Bioresources Utilization (Dalian Minzu University), Ministry of Education, Dalian 116600, P. R. China, ³Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea, ⁴Institute of Life Science and Natural Resources, Korea University, Seoul 02841, Republic of Korea¹Department of Bioengineering, College of Life Science, Dalian Minzu University, Dalian 116600, P. R. China, ²Key Laboratory of Biotechnology and Bioresources Utilization (Dalian Minzu University), Ministry of Education, Dalian 116600, P. R. China, ³Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea, ⁴Institute of Life Science and Natural Resources, Korea University, Seoul 02841, Republic of Korea

Corresponding author: Ki Hyun Nam , Tel: +82-10-5208-5730,

Received: 13 June 2018 • Revised: 16 July 2018 • Accepted: 18 July 2018

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Abstract

The Keima family comprises large Stokes shifts fluorescent proteins, which are useful for dual-color fluorescence crosscorrelation spectroscopy and multicolor imaging. dKeima570 belongs to the Keima family. It has a unique chromophore sequence composed of CYG with an emission peak at 570 nm, but its molecular properties are unclear. We report the spectral analysis of dKeima570 and its crystal structure at 2.0 Å resolution. The dKeima570 chromophore is mainly in the protonation state in the entire pH range. The pH-induced non-fluorescence state was observed below pH 4.0. The crystal structure of the dKeima570 chromophore has a cis conformation at pH 6.5. The chromophore is surrounded by a unique hydrogen bonding network containing a water bridge between Glu212 and Arg194. The analysis of the dimeric interface of dKeima570 revealed the key residues that maintain the oligomerization of Keima family. Structural comparisons of dKeima570 and mKeima provided insights into the unique large Stokes shifts characteristics of the Keima family.

Keywords: dKeima570; Keima; large Stokes shift; fluorescent protein; dimer interface

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Spectral and structural analysis of large Stokes shift fluorescent protein dKeima570

J. Microbiol. 2018;56(11):822-827. Published online October 24, 2018

DOI: https://doi.org/10.1007/s12275-018-8319-5

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