Near-infrared light activated protein photoswitches

近红外光激活蛋白质光开关

• 批准号: 8286092
• 负责人: Mark Gomelsky
• 金额: $ 18.21万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8286092
• 关键词: Address; Adenylate Cyclase; Adhesions; Affect; Animal Model; Animals; Apoptosis; Biological Process; Blood Glucose; Cardiac; Cardiovascular Physiology; Caspase; Cell Differentiation process; Cell Survival; Cell physiology; Cells; Cellular biology; Chemicals; Cyclic AMP; Cyclic GMP; Cyclic Nucleotides; Cysteine Protease; Data; Development; Developmental Biology; Diabetes Mellitus; Disease; Drosophila genus; Drosophila melanogaster; Engineering; Enzymes; Escherichia coli; Evaluation; Generations; Genes; Genetic; Genetic Screening; Government; Growth; Guanylate Cyclase; Homodimerization; Image; Immunology; In Vitro; Knowledge; Learning; Life; Light; Memory; Mission; Modeling; Mus; Neurobiology; Oncogenes; Output; Penetration; Persons; Pharmacologic Substance; Phosphotransferases; Photoreceptors; Phytochrome; Procedures; Process; Property; Proteins; Public Health; Recombinants; Research; Research Personnel; Resolution; Rhodobacter sphaeroides; Second Messenger Systems; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Source; Specificity; Stimulus; Study models; Testing; Tetrapyrroles; Time; Tissues; Transplantation; Variant; Visible Radiation; animal tissue; base; cancer cell; cell growth; cell killing; design; follow-up; gene therapy; in vivo; inhibitor/antagonist; innovation; insight; interdisciplinary approach; killings; model development; monomer; nervous system disorder; optogenetics; photoactivation; pro-caspase-3; protein function; research study; second messenger; small molecule; spatiotemporal; tool; tumor

DESCRIPTION (provided by applicant): The following contains proprietary/privileged information that M. Gomelsky requests not to be released to persons outside the Government, except for purposes of review and evaluation. PROJECT SUMMARY: Small molecule activators and inhibitors of signal transduction pathways are biologically useful, but are limited by their target specificities and spatiotemporal resolutio in vivo. A recently emerged optogenetic strategy can supplement chemical/pharmacological approaches. Ontogenetic involves introduction into cells and animals of genes encoding proteins whose activities can be photoactivated. Light is a unique stimulus in that it can control protein activities in vivo in a reversible manner and with spatiotemporal precision unattainable by chemicals. Photoreceptors of the bacteriophytochrome type absorb near-infrared light, which has superior tissue penetration properties, thus allowing protein photoactivation from unobtrusive external light sources. This is particularly important for studies on whole animals, such as mice. Recently, progress has been made in "transplanting" natural photoreceptor modules to control heterologous protein activities. Our long-term objective is to elucidate principles of engineering near-infrared light activated proteins using photosensory modules of bacteriophytochromes. This exploratory proposal will test the hypothesis that bacteriophytochrome photoreceptor domains can activate diverse homodimeric output activities. We will exploit our earlier studies of the BphG protein, a unique, near-infrared light activated diguanylyl cyclase. The nucleotidyl cyclases will be used as engineering targets. And an executioner (effectors) caspase rationally design bacteriophytochrome-based proteins, we will employ a multiprong approach involving circumventing our present inability to computational and structural analyses of proteins with genetic screening in E. coli. The proposed concept of engineering near-infrared light activated homodimeric proteins and the multidisciplinary approach to bacteriophytochrome engineering are innovative and feasible, as we already have constructed the first photoactivated homodimeric enzyme with a heterologous activity . Upon completion of this project, we anticipate to advance our understanding of the mechanism of light-induced signal propagation in bacteriophytochromes, and to uncover engineering principles for constructing homodimeric near-infrared light activated proteins. Because a large number of signaling proteins function as homodimers, light-induced protein homodimerization can be used to control a variety of cellular functions including apoptosis, differentiation, proliferation, transformation and adhesion. This research is significant because cAMP and cGMP control many cellular processes including growth, blood glucose levels, cardiac contractile function, and learning, memory and cancer cell survival. Photoactivated executioner caspase generated here will allow researchers to conduct targeted cell/tissue killing in whole animals using a mild and noninvasive procedure. These tools will likely find applications in cell biology, immunology and developmental biology, and potentially in cancer gene therapy. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because knowledge of functions and mechanisms of conserved signaling pathways is essential for our understanding of normal and disease states. Cyclic nucleotides, cAMP and cGMP, are universal second messengers that affect a variety of cellular functions. The ability to control second messengers in animal models with high spatial and temporal resolution has the potential to bring our understanding of tumorogenesis, cardiovascular function, development of diabetes, and neurological disorders to previously unattained levels. Caspases developed here will allow researchers to perform targeted cell killing in live animals using a mild and noninvasive treatment. Applications in cell biology, immunology and developmental biology, thus, the proposed research directly addresses the NIH's mission. And potentially in cancer gene therapy these tools will likely find.

描述（由申请人提供）：以下包含专有/特权信息，M. Gomelsky要求不向政府以外的人发布，除非用于审查和评估。