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The 4th ISGO International conference on Structural Genomics

作者： Joseph W.Arndt raymond c.stevens Department of Molecular Biology The Scripps Research Institute 10550 N.Torrey Pines RdLa JollaCA 92037USA Department of Molecular Biology The Scripps Research Institute 10550 N.Torrey Pines RdLa JollaCA 92037USA

＜正＞Botulinum neurotoxin is one of the most deadly molecules known and also a powerful protein therapeutic for neuromuscular disorders. Initially it was believed that there were only 7 different serotypes of botulinum neurotoxin. Recently, it has been discovered that there are more likely 50-200 different subtypes of botulinum

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USE OF STRUcTURAL PROTEOMIc METHODOLOGIES TOWARDS THE DEVELOPMENT OF PROTEIN THERAPEUTIcS

USE OF STRUCTURAL PROTEOMIC METHODOLOGIES TOWARDS THE DEVELO...

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The 4th ISGO International conference on Structural Genomics

作者： Alejandra Gámez Holly Archer raymond c.stevens Department of Molecular Biology The Scripps Research Institute 10550 N.Torrey Pines RdLa Jolla CA 92037 USA Department of Molecular Biology The Scripps Research Institute 10550 N.Torrey Pines RdLa Jolla CA 92037 USA Department of Molecular Biology The Scripps Research Institute 10550 N.Torrey Pines RdLa Jolla CA 92037 USA

＜正＞Employing the technologies and methodologies developed as part of the structural genomics efforts within the stevens laboratory at TSRI, we have developed a protein therapeutic for Phenylketonuria （PKU）. The disease PKU is an inherited metabolic disorder that results in phenylalanine accumulation in the body and can lead to irreversible brain damage. Initially, we analyzed all enzymes known to

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In vitro expression and analysis of the 826 human G protein-coupled receptors

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Protein & cell 2016年第5期7卷 325-337页

作者： Xuechen Lv Junlin Liu Qiaoyun Shi Qiwen Tan Dong Wu John J. Skinner Angela L. Walker Lixia Zhao Xiangxiang Gu Na chen Lu Xue Pei Si Lu Zhang Zeshi Wang Vsevolod Katritch Zhijie Liu raymond c. stevens iHuman Institute ShanghaiTech University Shanghai 201210 China Department of Biological Sciences Bridge Institute University of Southern California Los Angeles CA 90089 USA

G protein-coupled receptors （GPcRs） are involved in all human physiological systems where they are responsible for transducing extracellular signals into cells. GPcRs signal in response to a diverse array of stimuli including light, hormones, and lipids, where these signals affect downstream cascades to impact both health and disease states. Yet, despite their importance as therapeutic tar- gets, detailed molecular structures of only 30 GPcRs have been determined to date. A key challenge to their structure determination is adequate protein expression. Here we report the quantification of protein expression in an insect cell expression system for all 826 human GPcRs using two different fusion constructs. Expression char- acteristics are analyzed in aggregate and among each of the five distinct subfamilies. These data can be used to identify trends related to GPcR expression between dif- ferent fusion constructs and between different GPcR families, and to prioritize lead candidates for future structure determination feasibility.

关键词： G protein-coupled receptors insect,protein expression surface expression analysis fusionconstruct

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Structural basis for molecular recognition at serotonin receptors

Structural basis for molecular recognition at serotonin rece...

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第四届中国结构生物学学术讨论会

作者： Yi Jiang chong Wang Jinming Ma raymond c.stevens H.Eric Xu VARI/SIMM center CAS-Kay Laboratory of Receptor Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Department of Molecular Biology The Scripps Research Institute Laboratory of Structural Sciences Van Andel Research Institute

Serotonin or 5-hydroxytryptamine（5-HT） regulates a wide spectrum of human physiology through the 5-HT receptor family. We report the crystal structures of the human 5-HT1 B G protein-coupled receptor bound to the agonist anti-migraine medications ergotamine and dihydroergotamine. The structures reveal similar binding modes for these ligands, which occupy the orthosteric pocket and an extended binding pocket close to the extracellular loops. The orthosteric pocket is formed by residues conserved in the 5-HT receptor family, clarifying the family-wide agonist activity of 5-HT. compared to the accompanying structure of the 5-HT2 B receptor, the 5-HT1 B receptor displays a 3.0 angstrom outward shift at the extracellular end of helix V, resulting in a more open extended pocket that explains subtype selectivity. Together with docking and mutagenesis studies, these structures provide a comprehensive structural basis for understanding receptor-ligand interactions and designing subtype-selective serotonergic drugs.（Accepted by Science）

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Identification of natural products as novel ligands for the human 5-HT2c receptor

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Biophysics Reports 2018年第1期4卷 50-61页

作者： Yao Peng Simeng Zhao Yiran Wu Haijie cao Yueming Xu Xiaoyan Liu Wenqing Shui Jianjun cheng Suwen Zhao Ling Shen Jun Ma Ronald J.Quinn raymond c.stevens Guisheng Zhong Zhi-Jie Liu National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of Sciences Institute of Molecular and clinical Medicine Kunming Medical University iHuman Institute Shanghai Tech University University of chinese Academy of Sciences School of Life Science and Technology Shanghai Tech University college of Pharmacy Nankai University High-throughput Molecular Drug Discovery center Tianjin Joint Academy of Biotechnology and Medicine Eskitis Institute for Drug Discovery Griffith University

G protein-coupled receptors(GPcRs) constitute the largest human protein family with over 800 members,which are implicated in many important medical conditions. Serotonin receptors belong to the aminergic GPcR subfamily and play important roles in physiological and psychological activities. Structural biology studies have revealed the structures of many GPcRs in atomic details and provide the basis for the identification and investigation of the potential ligands, which interact with and modulate the ***, an integrative approach combining a focused target-specific natural compound library, a thermalshift-based screening method, affinity mass spectrometry, molecular docking, and in vitro as well as in vivo functional assay, was applied to identify(–)-crebanine and several other aporphine alkaloids as initial hits for a human serotonin receptor subtype, the 5-HT2creceptor. Further studies illuminated key features of their binding affinity, downstream signaling and tissue reaction, providing a molecular explanation for the interaction between(–)-crebanine and human 5-HT2creceptor.

关键词： GPcR 5-HT 2c receptor Natural product Alkaloids

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The JcIMPT - Innovative New Technologies for Membrane Protein Structural Biology

The JCIMPT - Innovative New Technologies for Membrane Protei...

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The 4th ISGO International conference on Structural Genomics

作者： raymond c.stevens Peter Kuhn Geoffrey chang Kurt Wüthrich Mark Yeager M.G.Finn Department of Molecular Biology10550 North Torrey Pines Road La JollaCA 92037USA Department of Chemistry The Scripps Research Institute 10550 North Torrey Pines RoadLa JollaCA 92037USA Department of cell Biology 10550 North Torrey Pines Road La JollaCA 92037USA Department of Molecular Biology 10550 North Torrey Pines Road La JollaCA 92037USA Department of Molecular Biology 10550 North Torrey Pines Road La JollaCA 92037USA Department of Chemistry The Scripps Research Institute10550 North Torrey Pines RoadLa JollaCA 92037USA Department of cell Biology 10550 North Torrey Pines Road La JollaCA 92037 USA Department of chemistry The Scripps Research Institute 10550 North Torrey Pines Road La JollaCA 92037 USA

＜正＞The mission of the Joint center for Innovative Membrane Protein Technologies （JcIMPT;http://***） is to develop and disseminate methods and technologies for structure-grade production and stabilization of integral membrane proteins. Miniaturization and automation are major components in the systematic investigation of membrane proteins and in the development of novel genetically engineered expression systems and sample preparation technologies.

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Structural Studies of HIV-1 co-receptor

Structural Studies of HIV-1 Co-receptor

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第一届国际暨第十三次中国生物物理学术大会

作者： Beili Wu Qiuxiang Tan Ya Zhu Gye Won Han Gustavo Fenalti Vsevolod Katritch Tracy Handel Vadim cherezov Qiang Zhao raymond c.stevens cAS Key Laboratory of Receptor Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Department of Molecule Biology The Scripps Research Institute 10550 North Torrey Pines Road La Jolla CA 92037 USA University of california San Diego Skaggs School of Pharmacy and Pharmaceutical Sciences La Jolla CA 92093 USA

＜正＞chemokine receptors are critical regulators of cell migration in the context of immune surveillance,inflammation and *** G protein-coupled chemokine receptors,cXcR4 and ccR5,are principle co-receptors for HIV-1 *** we report five independent crystal structures of cXcR4 bound to an antagonist small molecule IT1t and a

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用于膜蛋白研究的新型β-sheet多肽去垢剂

用于膜蛋白研究的新型β-sheet多肽去垢剂

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第八届全国化学生物学学术会议

作者：陶厚朝 Sung chang Lee Arne Moeller Fai Yiu Siu raymond c stevens clinton S Potter Bridget carragher Qinghai Zhang Department of Integrative Structural and computational Biology The Scripps Research InstituteLa JollaCaliforniaUSA The National Resource for Automated Molecular Microscopy The Scripps Research InstituteLa JollaCaliforniaUSA

＜正＞膜蛋白具有重要的生理功能,包括物质传递、能量转换和信号传导等。膜蛋白的研究对于生物医药领域的发展起着至关重要的作用,在FDA批准的上市药物中,目前约有一半以膜蛋白作为靶点。但是,限于研究手段,膜蛋白研究还很困难,远远落... 详细信息

＜正＞膜蛋白具有重要的生理功能,包括物质传递、能量转换和信号传导等。膜蛋白的研究对于生物医药领域的发展起着至关重要的作用,在FDA批准的上市药物中,目前约有一半以膜蛋白作为靶点。但是,限于研究手段,膜蛋白研究还很困难,远远落后于水溶性蛋白。膜蛋白的疏水性决定了其必须依靠合适的去垢剂才能稳定地存在于水溶液中。我们设计了一类基于β-strand/sheet的新型多肽

关键词：膜蛋白去垢剂稳定性电镜

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发展亲和质谱技术高通量筛选G-蛋白偶联受体的调节剂

发展亲和质谱技术高通量筛选G-蛋白偶联受体的调节剂

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第三届全国质谱分析学术报告会

作者：秦姗姗孟萌萌杨德华陆燕刘志杰王明伟 raymond c.stevens 水雯箐上海科技大学iHuman研究所

G-蛋白偶联受体（GPcR）家族是细胞表面最主要的受体,它们在各种病理生理条件下都起着核心作用[1]。许多研究都致力于发现高效率高选择性的GPcR新型受体,但是传统方法由于各种局限性而限制了相关的应用。近年来,表面离子共振技术（SPR... 详细信息

G-蛋白偶联受体（GPcR）家族是细胞表面最主要的受体,它们在各种病理生理条件下都起着核心作用[1]。许多研究都致力于发现高效率高选择性的GPcR新型受体,但是传统方法由于各种局限性而限制了相关的应用。近年来,表面离子共振技术（SPR）以及核磁共振技术（NMR）不断应用于GPcR配体的鉴定[2,3],但是需要高度纯化和比较稳定的蛋白,这对于一些GPcR来说是很难实现的,并且其通量较低,不适用于大规模的化合物库筛选。因此建立更加高效的配体筛选的方法尤为重要。为了解决这一问题,我们开发了基于亲和质谱方法的高通量、无标记的配体筛选方法用于GPcR小分子配体的筛选[4,5]。这种新方法可以用于筛选细胞表面GPcR的正构和异构配体。我们首先将它应用于5-羟色胺2c受体（5-HT2cR）的配体筛选中,5-HT2cR是用于治疗抑郁症,精神分裂症和其他精神障碍的抗精神病药物靶点。实验中我们共筛选了4333个化合物,鉴定到12个已知的5HT2cR的小分子调控剂,并发现若干未知的新结构配体。通过进一步的放射性同位素配体实验和细胞钙流实验,我们发现了一个作用于5HT2cR的新结构拮抗剂（Ic50≈1μM）。我们进而将这一亲和质谱新技术应用到另一个更具挑战性的GPcR靶标——-人胰高血糖素样肽-1受体（GLP-1R）。该受体是B类GPcR,它能介导肽激素GLP-1的作用,并且在葡萄糖的体内平衡中发挥着重要的作用,是公认的糖尿病药物研发的重要靶标蛋白。但是对于该受体,目前已报道的非肽类的小分子调节剂少之又少。利用我们的高通量亲和质谱筛选技术,结合放射性同位素配体实验以及胞内信号转导实验,我们发现了4个Ec50在微摩尔级的新型别构激动剂。综上所述,我们所发展的膜受体亲和质谱筛选技术主要有以下优点:1)蛋白和小分子都是无标记的;2)对细胞膜上表达的野生型受体直接进行筛选,保证相互作用处于天然状态;3)直接检测蛋白质和小分子相互作用,有助于发现靶蛋白的别构调节剂;4)该实验不需要纯化膜蛋白,从而大大降低了实验成本,避免了膜蛋白纯化的固有局限。

关键词： G-蛋白偶联受体亲和质谱高通量 5-HT2cR GLP-1R

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Dynamics of self-hybridized exciton–polaritons in 2D halide perovskites

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Light(Science & Applications) 2024年第1期13卷 13-22页

作者： Surendra B.Anantharaman Jason Lynch christopher E.stevens christopher Munley chentao Li Jin Hou Hao Zhang Andrew Torma Thomas Darlington Francis coen Kevin Li Arka Majumdar PJames Schuck Aditya Mohite Hayk Harutyunyan Joshua R.Hendrickson Deep Jariwala Department of Electrical and Systems Engineering University of PennsylvaniaPhiladelphiaPA19104USA KBR Inc. BeavercreekOH45431USA Air Force Research Laboratory Sensors DirectorateWright-Patterson Air Force BaseOH45433USA Department of Physics University of WashingtonSeattleWA98195USA Department of Physics Emory UniversityAtlantaGA30322USA Department of chemical and Biomolecular Engineering Rice UniversityHoustonTX77005USA Department of Materials Science and Nanoengineering Rice UniversityHoustonTX77005USA Applied Physics Program Smalley‐Curl InstituteRice UniversityHoustonTX77005USA Department of Mechanical Engineering Columbia UniversityNew YorkNY10027USA Department of Electrical and computer Engineering University of WashingtonSeattleWA98195USA

Excitons,bound electron–hole pairs,in two-dimensional hybrid organic inorganic perovskites(2D HOIPs)are capable of forming hybrid light-matter states known as exciton-polaritons(E–Ps)when the excitonic medium is confined in an optical *** the case of 2D HOIPs,they can self-hybridize into E–Ps at specific thicknesses of the HOIP crystals that form a resonant optical cavity with the ***,the fundamental properties of these self-hybridized E–Ps in 2D HOIPs,including their role in ultrafast energy and/or charge transfer at interfaces,remain ***,we demonstrate that>0.5µm thick 2D HOIP crystals on Au substrates are capable of supporting multiple-orders of self-hybridized E–P *** E–Ps have high Q factors(>100)and modulate the optical dispersion for the crystal to enhance sub-gap absorption and *** varying excitation energy and ultrafast measurements,we also confirm energy transfer from higher energy E–Ps to lower energy E–***,we also demonstrate that E–Ps are capable of charge transport and transfer at *** findings provide new insights into charge and energy transfer in E–Ps opening new opportunities towards their manipulation for polaritonic devices.

关键词： charge polar excitation

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