Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction

doi:10.1107/S1399004713032434

KMS > iHuman研究所

	Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction
	Natashin, Pavel V.1,2,3; Ding, Wei 1,4; Eremeeva, Elena V.2,3; Markova, Svetlana V.2,3; Lee, John 5; Vysotski, Eugene S.2,3; Liu, Zhi-Jie1,6
	2014-03
发表期刊	ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY (IF:7.232[JCR-2021],9.416[5-Year])
ISSN	1399-0047
卷号	70 页码:720-732
发表状态	已发表
DOI	10.1107/S1399004713032434
摘要	Ca2+-regulated photoproteins, which are responsible for light emission in a variety of marine coelenterates, are a highly valuable tool for measuring Ca2+ inside living cells. All of the photoproteins are a single-chain polypeptide to which a 2-hydroperoxycoelenterazine molecule is tightly but noncovalently bound. Bioluminescence results from the oxidative decarboxylation of 2-hydroperoxycoelenterazine, generating protein-bound coelenteramide in an excited state. Here, the crystal structures of the Y138F obelin mutant before and after bioluminescence are reported at 1.72 and 1.30 angstrom resolution, respectively. The comparison of the spatial structures of the conformational states of Y138F obelin with those of wild-type obelin gives clear evidence that the substitution of Tyr by Phe does not affect the overall structure of both Y138F obelin and its product following Ca2+ discharge compared with the corresponding conformational states of wild-type obelin. Despite the similarity of the overall structures and internal cavities of Y138F and wild-type obelins, there is a substantial difference: in the cavity of Y138F obelin a water molecule corresponding to W2 in wild-type obelin is not found. However, in Ca2+-discharged Y138F obelin this water molecule now appears in the same location. This finding, together with the observed much slower kinetics of Y138F obelin, clearly supports the hypothesis that the function of a water molecule in this location is to catalyze the 2-hydroperoxycoelenterazine decarboxylation reaction by protonation of a dioxetanone anion before its decomposition into the excited-state product. Although obelin differs from other hydromedusan Ca2+-regulated photoproteins in some of its properties, they are believed to share a common mechanism.
收录类别	SCI
语种	英语
资助项目	Russian Federation 'Leading Science School'[3951.2012.4]
WOS研究方向	Biochemistry & Molecular Biology ; Biophysics ; Crystallography
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biophysics ; Crystallography
WOS记录号	WOS:000332406600011
出版者	WILEY-BLACKWELL
WOS关键词	AEQUORIN BIOLUMINESCENCE ; SEQUENCE-ANALYSIS ; CRYSTAL-STRUCTURE ; CA2+-BINDING PHOTOPROTEIN ; VIOLET BIOLUMINESCENCE ; CALCIUM CONCENTRATION ; ANGSTROM RESOLUTION ; RECOMBINANT OBELIN ; MNEMIOPSIS-LEIDYI ; EXCITED-STATES
原始文献类型	Article
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/1072
专题	iHuman研究所 iHuman研究所_PI研究组_刘志杰组
通讯作者	Vysotski, Eugene S.
作者单位	1.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100080, Peoples R China 2.Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia 3.Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Lab Bioluminescence Biotechnol, Chair Biophys, Krasnoyarsk, Russia 4.Chinese Acad Sci, Inst Biophys, Ctr Biol Imaging, Beijing 100080, Peoples R China 5.Univ Georgia, Dept Biochem Mol Biol, Athens, GA 30602 USA 6.Shanghai Tech Univ, Human Inst, Shanghai, Peoples R China
推荐引用方式 GB/T 7714	Natashin, Pavel V.,Ding, Wei,Eremeeva, Elena V.,et al. Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction[J]. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY,2014,70:720-732.
APA	Natashin, Pavel V..,Ding, Wei.,Eremeeva, Elena V..,Markova, Svetlana V..,Lee, John.,...&Liu, Zhi-Jie.(2014).Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction.ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY,70,720-732.
MLA	Natashin, Pavel V.,et al."Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction".ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY 70(2014):720-732.