CAREER: Using Laboratory Evolution to Study Protein Dynamics In Vivo

职业：利用实验室进化研究体内蛋白质动力学

• 批准号: 1150138
• 负责人: Jonathan Silberg
• 金额: $ 62.96万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1150138&HistoricalAwards=false
• 关键词: CAREER; Using; Laboratory; Evolution; Study

Intellectual MeritIn order to better understand which atoms in a protein contribute decisively to the key properties that support enzyme function, this project will generate libraries of natural proteins that contain different classes of mutations (random substitutions, glycine substitutions, backbone fission, circular permutation, insertions, and deletions) and examine how each mutation type affects protein folding and function within cells. Adenylate kinases will be used for these studies, since an abundance of pre-existing biophysical data can be tapped to interpret results from systematic mutagenesis experiments. This research will establish if the pattern of a protein's functional tolerance to different classes of mutation contains information on how different native positions within its primary sequence contribute to structure, stability, folding, dynamics, and function. This experimental strategy will be potentially useful for studying any protein for which an in vivo screen is available, including poorly behaved proteins that cannot readily be examined by in vitro physical methods. Broader ImpactThis broader impact of this project will be to provide training for graduate and undergraduate students interested in science careers. The PI and graduate students performing this research will conduct annual round table discussions for undergraduates enrolled at a nearby community college. These community college students begin their education in lecture courses and are not necessarily exposed to an environment that mirrors a research lab. Round table discussions will be used to increase student self-confidence by showing beginning college students that they can read a peer-reviewed research article, demonstrate to these students the benefits of research training, and facilitate community college student recruitment to research experiences. These efforts will vertically integrate undergraduate and graduate education by providing beneficial training for graduate students, building self-sustaining outreach from a research university to a minority serving community college, and offering research experiences for community college students.

为了更好地了解蛋白质中哪些原子对支持酶功能的关键特性起决定性作用，该项目将生成包含不同类型突变（随机取代，甘氨酸取代，骨架分裂，环状排列，插入和缺失）的天然蛋白质库，并检查每种突变类型如何影响细胞内的蛋白质折叠和功能。这些研究将使用腺苷酸激酶，因为可以利用丰富的已有生物物理数据来解释系统性突变实验的结果。这项研究将确定蛋白质对不同类型突变的功能耐受性模式是否包含关于其一级序列内不同天然位置如何有助于结构，稳定性，折叠，动力学和功能的信息。这种实验策略将是潜在的有用的研究任何蛋白质的体内屏幕是可用的，包括表现不佳的蛋白质，不能很容易地通过体外物理方法进行检查。 更广泛的影响这个项目的更广泛的影响将是为对科学职业感兴趣的研究生和本科生提供培训。进行这项研究的PI和研究生将为附近社区大学的本科生进行年度圆桌讨论。这些社区学院的学生开始他们的教育在讲座课程，并不一定接触到一个环境，反映了一个研究实验室。圆桌讨论将被用来增加学生的自信心，通过展示开始大学生，他们可以阅读同行评议的研究文章，向这些学生展示研究培训的好处，并促进社区大学的学生招聘研究经验。这些努力将垂直整合本科和研究生教育，为研究生提供有益的培训，建立从研究型大学到少数服务社区学院的自我维持外联，并为社区学院学生提供研究经验。