Crystal structure of semisynthetic obelin-v

doi:10.1002/pro.4244

	Crystal structure of semisynthetic obelin-v
	Larionova, Marina D.1,2 ; Wu, Lijie2 ; Eremeeva, Elena, V 1,3; Natashin, Pavel, V 1; Gulnov, Dmitry, V 3; Nemtseva, Elena, V 1,3; Liu, Dongsheng2 ; Liu, Zhi-Jie2,4 ; Vysotski, Eugene S.1
	2022-02
发表期刊	PROTEIN SCIENCE (IF:4.5[JCR-2023],9.1[5-Year])
ISSN	0961-8368
EISSN	1469-896X
DOI	10.1002/pro.4244
摘要	Coelenterazine-v (CTZ-v), a synthetic derivative with an additional benzyl ring, yields a bright bioluminescence of Renilla luciferase and its yellow mutant with a significant shift in the emission spectrum toward longer wavelengths, which makes it the substrate of choice for deep tissue imaging. Although Ca2+-regulated photoproteins activated with CTZ-v also display red-shifted light emission, in contrast to Renilla luciferase their bioluminescence activities are very low, which makes photoproteins activated by CTZ-v unusable for calcium imaging. Here, we report the crystal structure of Ca2+-regulated photoprotein obelin with 2-hydroperoxycoelenterazine-v (obelin-v) at 1.80 angstrom resolution. The structures of obelin-v and obelin bound with native CTZ revealed almost no difference; only the minor rearrangement in hydrogen-bond pattern and slightly increased distances between key active site residues and some atoms of 2-hydroperoxycoelenterazine-v were found. The fluorescence quantum yield (phi(FL)) of obelin bound with coelenteramide-v (0.24) turned out to be even higher than that of obelin with native coelenteramide (0.19). Since both obelins are in effect the enzyme-substrate complexes containing the 2-hydroperoxy adduct of CTZ-v or CTZ, we reasonably assume the chemical reaction mechanisms and the yields of the reaction products (phi(R)) to be similar for both obelins. Based on these findings we suggest that low bioluminescence activity of obelin-v is caused by the low efficiency of generating an electronic excited state (phi(S)). In turn, the low phi(S) value as compared to that of native CTZ might be the result of small changes in the substrate microenvironment in the obelin-v active site.
关键词	analog bioluminescence coelenterazine coelenterazine-v obelin photoprotein protein structure
收录类别	SCIE
语种	英语
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000723224900001
出版者	WILEY
原始文献类型	Article; Early Access
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/133139
专题	iHuman研究所_PI研究组_刘志杰组生命科学与技术学院 iHuman研究所 iHuman研究所_特聘教授组_Kurt Wuthrich组 iHuman研究所_科学装置(X)_膜蛋白同步辐射线站
通讯作者	Liu, Zhi-Jie; Vysotski, Eugene S.
作者单位	1.SB RAS, Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Photobiol Lab, Inst Biophys, Krasnoyarsk, Russia; 2.ShanghaiTech Univ, iHuman Inst, Ren Bldg,393 Middle Huaxia Rd, Shanghai 201210, Peoples R China; 3.Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Krasnoyarsk, Russia; 4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
第一作者单位	iHuman研究所
通讯作者单位	iHuman研究所; 生命科学与技术学院
推荐引用方式 GB/T 7714	Larionova, Marina D.,Wu, Lijie,Eremeeva, Elena, V,et al. Crystal structure of semisynthetic obelin-v[J]. PROTEIN SCIENCE,2022.
APA	Larionova, Marina D..,Wu, Lijie.,Eremeeva, Elena, V.,Natashin, Pavel, V.,Gulnov, Dmitry, V.,...&Vysotski, Eugene S..(2022).Crystal structure of semisynthetic obelin-v.PROTEIN SCIENCE.
MLA	Larionova, Marina D.,et al."Crystal structure of semisynthetic obelin-v".PROTEIN SCIENCE (2022).