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LOV to BLUF： Flavoprotein Contributions to the Optogenetic Toolkit

Molecular Plant 2012年第3期5卷 533-544页

作者： John M. Christie Jayde Gawthorne Gillian Young Niall J. Fraser^c and Andrew J. Roe Institute of Molecular Cell and Systems Biology College of Medical Veterinary and Life Sciences University of Glasgow UK Institute of Infection immunity and Inflammation College of Medical Veterinary and Life Sciences University of Glasgow UK Institute of Cardiovascular and Medical Sciences College of Medical Veterinary and Life Sciences University of Glasgow UK

Optogenetics is an emerging field that combines optical and genetic approaches to non-invasively interfere with cellular events with exquisite spatiotemporal control. Although it arose originally from neuroscience, optogenetics is widely applicable to the study of many different biological systems and the range of applications arising from this tech- nology continues to increase. Moreover, the repertoire of light-sensitive proteins used for devising new optogenetic tools is rapidly expanding. Light, Oxygen, or Voltage sensing （LOV） and Blue-Light-Utilizing flavin adenine dinucleotide （FAD）（BLUF） domains represent new contributors to the optogenetic toolkit. These small （100-140-amino acids） flavoprotein modules are derived from plant and bacterial photoreceptors that respond to UV-A/blue light. In recent years, considerable progress has been made in uncovering the photoactivation mechanisms of both LOV and BLUF domains. This knowledge has been applied in the design of synthetic photoswitches and fluorescent reporters with applications in cell biology and biotechnology. In this review, we summarize the photochemical properties of LOV and BLUF photosensors and highlight some of the recent advances in how these flavoproteins are being employed to artificially regulate and image a variety of biological processes.

关键词： Blue-Light.Utilizing FAD chromophore flavin fluorescence light, oxygen or voltage optogenetics photo- receptor protein engineering

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Affinity-induced covalent protein-protein ligation via the SpyCatcherSpyTag interaction

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Green Carbon 2023年第1期1卷 33-42页

作者： Jacob O.Fierer Omar E.Tovar-Herrera Jonathan Y.Weinstein Amaranta Kahn Sarah Moraïs Itzhak Mizrahi Edward A.Bayer Department of Biomolecular Sciences the Weizmann Institute of ScienceRehovot 7610001Israel Department of Life Sciences Ben-Gurion University of the NegevBeer Sheva 84105Israel National Institute of Biotechnology in the Negev Ben-Gurion University of the NegevBe’er Sheva 84105Israel The Goldman Sonnenfeldt School of Sustainability and Climate Change Ben-Gurion University of the NegevBeer Sheva 84105Israel

Production of economically viable bioethanol is potentially an environmentally and financially worthwhile *** major source for fermentable sugars is ***,lignocellulosic biomass is difficult to degrade,owing to its inherent structural *** are complexes of cellulases and associated polysaccharide-degrading enzymes bound to a protein scaffold that can efficiently degrade *** of the enzyme subunits into the complex depends on intermodular cohesin-dockerin interactions,which are robust but nonetheless *** modular architecture of these complexes can be used to assemble artificial designer cellulosomes for advanced nanotechnological *** that promote lignocellulose degradation involve high temperatures and acidic or alkaline conditions that could dismember designer cellulosomes,thus requiring separation of reaction steps,thereby increasing overall process *** overcome these challenges,we developed a means of covalently locking cohesin-dockerin interactions by integrating the chemistry of SpyCatcher-SpyTag approach to target and secure the *** resultant cohesin-conjugated dockerin complex was resistant to high temperatures,SDS,and urea while high affinity and specificity of the interacting modular components were *** this approach,a covalently locked,bivalent designer cellulosome complex was produced and demonstrated to be enzymatically active on cellulosic *** combination of affinity systems with SpyCatcher-SpyTag chemistry may prove of general use for improving other types of protein ligation systems and creating unconventional,biologically active,covalently locked,affinity-based molecular architectures.

关键词： Synthetic Biology protein Ligation Cellulosomes Cohesin Dockerin protein engineering Cellulose Degradation

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Mechanism of multi-point site-directed mutagenesis

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Progress in Natural Science:Materials International 1997年第3期 48-55页

作者：谢毅肖谷田林卿王顺德叶韫辉吴小舟

Subtilisin E gene was mutagemzed in vitro as a model in one-round mutagenesis with multiple chemical-synthesized mutagenic oligonucleotides. The rationale of the new method is suggested The elements that influence the mutant efficiency such as the type, amount, distribution and mutual location of primers, and the primer to template molar ratio are discussed

关键词： site-directed mutagenesis multi-point site-specific mutagenesis protein engineering subtilisin E.

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A novel fluorescent sensor for cAMP based on *** CAP protein

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Journal of Chinese Pharmaceutical Sciences 2019年第8期28卷 556-560页

作者： Jiayuan Zhang Yuxin Song Chuchen Wang Jing Wang State Key Laboratory of Natural and Biomimetic Drugs Department of Chemical BiologySchool of Pharmaceutical Sciences Peking University Health Science Center Beijing 100191 China

cAMP is an important second messenger that is capable of controlling a wide array of cellular processes, including glycogen, sugar and lipid metabolism. Here we report the design and construction of a novel genetically encoded fluorescent sensor for cAMP. The sensor was realized by fusing E. coli CAP protein with cpYFP, and displayed a one-fold fluorescence change towards c AMP binding. Further characterization assays demonstrated that the sensor had high affinity for cAMP and fast response *** development of our sensor could be a useful supplement to existing methods for cAMP detection.

关键词： c AMP Fluorescent sensor protein engineering Signal transduction

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A NEW INTERFERON α1 MUTANT(HuIFN-α1/86D)POSSESSES HIGHER BIOLOGICAL ACTIVITIES AND THERMAL STABILITY

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Progress in Natural Science:Materials International 1991年第1期 65-67页

作者：候云德王伟李燕 Chinese Academy of Preventive Medicine.Beijing 100052 National Laboratory of Molecular Virology and Genetic engineering Institute of Virology PRC

Human x interferon genomes are divided into two groups:IFN-α1 and IFN-α***-x1 group,there are at least 15 members,which are somewhat different in their molecu-lar structures and *** 166 amino acids,about half of the... 详细信息

关键词： interferon mutant antiviral protein protein engineering

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Cellular Synthesis of Topological proteins

Cellular Synthesis of Topological Proteins

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第十一届全国化学生物学学术会议

作者：张文彬北京大学

Chemical topology has emerged as a unique dimension for protein *** has demonstrated the power of protein topology engineering in a small yet elegant set of proteins with nontrivial topology（such as cyclotides and lasso peptides possessing exceptional bioactivity and stability）.However,artificial topological proteins remain *** this talk,I will discuss our recent efforts in the field of protein topology engineering from the development of genetically encoded protein chemical tools to the cellular synthesis of topological/mechano-proteins and further to the their biological significance and potential *** strategies such as the "assembly-reaction" synergy and the active-template method,we have prepared various topological proteins including cyclic proteins,star proteins,protein catenanes,lasso proteins,protein heterocatenanes and *** unique topologies have been proven by combined techniques such as controlled digestion experiments,NMR spectrometry,and protein *** them,proteins with mechanical bonds（"mechanoproteins"） are of particular interest owing to their potential functional benefits such as structure stabilization,quaternary structure control,synergistic multivalency effect and dynamic mechanical sliding/switching ***,protein catenanes containing folded structural domains have been found to exhibit enhanced stability toward proteolytic digestion,heat/chemical denaturation,and freezethawing treatments,which are highly desired features for enzymes in industry and protein *** results suggest that topology is indeed a unique dimension for protein engineering

关键词： protein engineering topology catenane SpyStapler SpyTag

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Modeling and predicting the activities of trans-acting splicing factors with machining learning

Modeling and predicting the activities of trans-acting splic...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2018年全国学术会议

作者： Miaowei Mao Yue Hu Yun Yang Yajie Qian Huanhuan Wei Wei Fan Yi Yang Xiaoling Li Zefeng Wang CAS Key Laboratory of Computational Biology CAS-MPG Partner Institute for Computational BiologyShanghai Institute of Nutrition and HealthShanghai Institutes for Biological SciencesUniversity of Chinese Academy of SciencesCAS Center for Excellence in Molecular Cell ScienceChinese Academy of Sciences Synthetic Biology and Biotechnology Laboratory State Key Laboratory of Bioreactor EngineeringSchool of PharmacyEast China University of Science and Technology Signal Transduction Laboratory National Institute of Environmental Health Sciences

Alternative splicing(AS) is generally regulated by trans-splicing factors that specifically bind to cis-elements in *** genome encodes ～1500 RNA binding proteins(RBPs) that potentially regulate AS,yet their function remain largely *** explore their potential activities,we fused the putative functional domains of RBPs to a sequence-specific RNAbinding domain,and systemically analyzed how these engineered factors affect *** discovered that ～80% of low complexity domains in endogenous RBPs displayed distinct context-dependent activities in regulating splicing,indicating that AS is under more extensive regulation than previously *** developed a machine learning approach classify and predict the activities of RBPs based on their sequence compositions,and further validated this model using endogenous RBPs and synthetic *** results represent a systematic inspection,modeling,prediction and validation of how RBP sequences affect their activities in controlling splicing,paving the way for de novo engineering of artificial splicing factors.

关键词： Alternative splicing Splicing factors Machine learning RNA binding domains protein engineering protein activity prediction

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原位定点可控合成高分子-蛋白质结合体及其在生物医药中的应用

原位定点可控合成高分子-蛋白质结合体及其在生物医药中的应用

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2012年全国高分子材料科学与工程研讨会

作者：高卫平清华大学医学院生物医学工程系

＜正＞蛋白质是具有生物活性和特异性的生物大分子,己作为高端药物（如抗体、Exendin-4和干扰素等）在临床上使用,并且正在成为医药市场的主流药物。尽管如此,蛋白质的一个主要缺陷是体内循环半衰期较短,药物代谢动力学差。目前,提高蛋... 详细信息

＜正＞蛋白质是具有生物活性和特异性的生物大分子,己作为高端药物（如抗体、Exendin-4和干扰素等）在临床上使用,并且正在成为医药市场的主流药物。尽管如此,蛋白质的一个主要缺陷是体内循环半衰期较短,药物代谢动力学差。目前,提高蛋白质半衰期的一个有效

关键词： Polymer-protein conjugate polymer engineering protein engineering drug delivery

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