Structural Determinants for Light-Dependent Membrane Binding of a Photoswitchable Polybasic Domain

doi:10.1021/acssynbio.0c00571

	Structural Determinants for Light-Dependent Membrane Binding of a Photoswitchable Polybasic Domain
	Li, Ling 1,2; He, Lian 3; Wu, Bo 1; Yu, Chuandi 1,2; Zhao, Hongxin 1; Zhou, Yubin 3; Wang, Junfeng 1,2,4; Zhu, Lei 1
刊名	ACS SYNTHETIC BIOLOGY
	2021-03-19
卷号	10
关键词	polybasic domain LOV2 optogenetics NMR PI(4,5)P-2 protein-lipid interaction
ISSN号	2161-5063
DOI	10.1021/acssynbio.0c00571
通讯作者	Zhou, Yubin(yubinzhou@tamu.edu) ; Wang, Junfeng(junfeng@hmfl.ac.cn) ; Zhu, Lei(zhulei@hmfl.ac.cn)
英文摘要	OptoPB is an optogenetic tool engineered by fusion of the phosphoinositide (PI)-binding polybasic domain of Ritl (Rit-PB) to a photoreactive light-oxygen-voltage (LOV) domain. OptoPB selectively and reversibly binds the plasma membrane (PM) under blue light excitation, and in the dark, it releases back to the cytoplasm. However, the molecular mechanism of optical r egulation and lipid recognition is still unclear. Here using nuclear magnetic resonance (NMR) spectroscopy, liposome pulldown assay, and surface plasmon resonance (SPR), we find that OptoPB binds to membrane mimetics containing di- or triphosphorylated phosphatidylinositols, particularly phosphatidylinositol 4,5-bisphosphate (PI(4,5)P-2), an acidic phospholipid predominantly located in the eukaryotic PM. In the dark, steric hindrance prevented this protein-membrane interaction, while 470 nm blue light illumination activated it. NMR titration and site-directed mutagenesis revealed that both cationic and hydrophobic Rit-PB residues are essential to the membrane interaction, indicating that OptoPB binds the membrane via a specific PI(4,5)P-2-dependent mechanism.
资助项目	National Natural Science Foundation of China[31800645] ; National Natural Science Foundation of China[U1532269] ; National Natural Science Foundation of China[21673244] ; Anhui Provincial Natural Science Foundation[18085QC65] ; Welch Foundation[BE-1913-20190330] ; American Cancer Society[RSG-16-215-01-TBE] ; High Magnetic Field Laboratory of Anhui Province
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000631444600011
资助机构	National Natural Science Foundation of China ; Anhui Provincial Natural Science Foundation ; Welch Foundation ; American Cancer Society ; High Magnetic Field Laboratory of Anhui Province
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/120804]
专题	中国科学院合肥物质科学研究院
通讯作者	Zhou, Yubin; Wang, Junfeng; Zhu, Lei
作者单位	1.Chinese Acad Sci, Hefei Inst Phys Sci, High Magnet Field Lab, CAS Key Lab High Magnet Field & Ion Beam Phys Bio, Hefei 230031, Peoples R China 2.Univ Sci & Technol China, Hefei 230026, Peoples R China 3.Texas A&M Univ, Ctr Translat Canc Res, Inst Biosci & Technol, Houston, TX 77030 USA 4.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei 230031, Peoples R China
推荐引用方式 GB/T 7714	Li, Ling,He, Lian,Wu, Bo,et al. Structural Determinants for Light-Dependent Membrane Binding of a Photoswitchable Polybasic Domain[J]. ACS SYNTHETIC BIOLOGY,2021,10.
APA	Li, Ling.,He, Lian.,Wu, Bo.,Yu, Chuandi.,Zhao, Hongxin.,...&Zhu, Lei.(2021).Structural Determinants for Light-Dependent Membrane Binding of a Photoswitchable Polybasic Domain.ACS SYNTHETIC BIOLOGY,10.
MLA	Li, Ling,et al."Structural Determinants for Light-Dependent Membrane Binding of a Photoswitchable Polybasic Domain".ACS SYNTHETIC BIOLOGY 10(2021).