RUI: Thermochemistry, Reaction Dynamics and Conformational Changes that Accompany the Collisional Activation of Peptide Ternary Complexes and Recombinant Tagged Proteins

在化学结构、动力学机制-化学部B计划的支持下，德克萨斯州A M大学商务部的Laurence Angel和Venugopalan Cheriyath将使用电喷雾电离-离子迁移-质谱技术开发新的锌和镍结合小肽。在这项研究中表征和优化的肽可能会导致改进的治疗性蛋白质纯化的发展。主要的教育目标是吸引，鼓励和留住在科学中代表性不足的学生;特别是在化学，生物学和物理学的关键交叉点。参与研究的本科生将获得尖端仪器，生物化学和理论技术的知识和经验，并在科学会议和同行评审的期刊出版物上分享他们的发现。该研究将确定替代金属结合（AMB）肽的pH依赖性反应性和金属离子亲和力。将通过电喷雾电离-离子迁移率-质谱（ESI-IM-MS）技术测量pH依赖性反应性。金属离子亲和力将从阈值碰撞诱导解离（TCID）分析确定，用于确定三元络合物的解离反应的热化学。TCID分析需要反应物和产物的真实分子参数，我们将开发结合分子动力学模拟和PM 7的新程序，用于定位具有与ESI-IM-MS测量的碰撞截面一致的最低能量构象。研究将检查（i）[ambm+M（II）+ambn]和[ambm+M（II）+NTA]三元复合物的解离反应的热化学，其中M = Ni或Zn和NTA =次氮基三乙酸;（ii）与三元络合物的不同构象相关的pH依赖性热化学;（iii）三元络合物解离的分子动力学模拟;（iv）选择的AMB作为用于表达和纯化重组蛋白的标签，和（v）使用碰撞分析的AMB标记的蛋白构象稳定性。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Structure, Dynamics & Mechanisms-B Program of the Chemistry Division, Laurence Angel and Venugopalan Cheriyath of Texas A&M University - Commerce will develop new zinc- and nickel-binding small peptides using electrospray ionization - ion mobility - mass spectrometry techniques. Peptides characterized and optimized in this study may lead to the development of improved therapeutic protein purification. The primary educational goal is to engage, encourage, and retain students underrepresented in science; specifically, at the critical intersection of chemistry, biology and physics. Undergraduate students involved in the research will gain knowledge and experience of cutting edge instrumental, biochemical and theoretical techniques and share their findings at scientific meetings and in peer-reviewed journal publications. The study will determine the pH-dependent reactivity and metal ion affinity of alternative metal binding (AMB) peptides. Measurement of the pH-dependent reactivity will be done by electrospray ionization - ion mobility - mass spectrometry (ESI-IM-MS) techniques. The metal ion affinities will be determined from threshold collision-induced dissociation (TCID) analyses for determining the thermochemistry of the dissociation reactions of ternary complexes. TCID analyses requires realistic molecular parameters of the reactants and products and we will develop new procedures combining molecular dynamic simulations and PM7 for locating the lowest-energy conformers with collision cross sections that agree with those measured by ESI-IM-MS. The research will examine (i) thermochemistry of the dissociation reactions of [ambm+M(II)+ambn] and [ambm+M(II)+NTA] ternary complexes, where M = Ni or Zn and NTA = nitrilotriacetic acid; (ii) pH-dependent thermochemistry relating to different conformers of the ternary complexes; (iii) molecular dynamics simulations of the dissociation of the ternary complexes; (iv) selected AMBs as tags for the expression and purification of recombinant protein and (v) AMB-tagged protein conformational stability using collision-induced unfolding analyses.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.