CAA: Mapping the Topology of Neuronal Nitric Oxide Synthase Bound to Calmodulin

摘要一氧化氮合酶是一种参与神经细胞信号传导和血管张力等多种生物学功能的酶。这种酶以三种不同的形式存在，在体内受到不同的调节。其中一个调节因素是这种酶与另一种叫做钙调蛋白的蛋白质的相互作用。钙调素（CaM）实际上参与调节细胞中40多种不同的酶，其作用尚未完全了解。这里概述的项目将利用传统的蛋白质足迹方法结合质谱来揭示CaM结合和调节一氧化氮合酶（nNOS）神经元形式的机制。更具体地说，这项研究将深入了解蛋白质的折叠结构，因为它们与神经元细胞的功能有关。这项研究的更广泛的影响是，它将提供关于CaM在调节细胞功能中的作用的急需信息，特别是将阐明nNOS的调节。这项研究还将为卡拉马祖学院的本科生提供在基因组学和蛋白质组学的前沿进行研究的机会。通过将质谱研究与生物信息学技术相结合，毕业生将更好地准备在生物化学研究的新时代中前进。这项研究将增加霍华德休斯医学研究所对学院生物化学和分子生物学项目的承诺，以支持将生物信息学模块纳入教学课程。这项研究的资金还将帮助PI，一个未被充分代表的少数民族的成员，吸引和留住未被充分代表的生化研究学生，并将为这些学生提供一个具有挑战性和令人兴奋的问题来研究。卡拉马祖学院最近获得了一项NSF S-STEM资助，用于向学院新招收的代表性不足的学生提供奖学金，这些学生计划将科学和数学作为一门学习课程。PI计划使用获得的资金为S-STEM学者提供夏季津贴，以便他们最早在第一年的夏季参与研究项目。

ABSTRACTNitric oxide synthase is an enzyme involved in a number of biological functions such as neuronal cell signaling and vascular tone. This enzyme exists as three distinct forms that are regulated differently in the body. One regulatory factor is the enzyme''s interaction with another protein called calmodulin. Calmodulin (CaM) is in fact involved in regulating over 40 different enzymes in cells, an action not completely understood. The project outlined here will make use of traditional protein footprinting methodologies coupled with mass spectrometry to unravel the mechanisms underlying CaM''s binding to and regulating the neuronal form of nitric oxide synthase (nNOS). More specifically, this research will give insights into the folded structure of the proteins as they relate to the functioning of neuronal cells. The broader impacts of this research are that it will provide much needed information regarding the role of CaM in regulating cellular functions, and in particular will shed light on the regulation of nNOS. This research will additionally provide undergraduate students at Kalamazoo College the opportunity to conduct research at the cutting-edge of genomics and proteomics. By conducting mass spectrometric studies coupled with bioinformatics techniques, graduates will be better prepared to advance in this new era of biochemistry research. This research will add to the commitment to the biochemistry and molecular biology programs at the College already made by the Howard Hughes Medical Institute to support bioinformatics modules incorporated into teaching curriculum. Funding for this research will additionally help the PI, a member of an underrepresented minority, attract and retain underrepresented students in biochemical research, and will provide these students a challenging and exciting problem to study. Kalamazoo College has been recently awarded an NSF S-STEM grant to be used to provide scholarships to newly recruited underrepresented students to the College who plan to pursue science and math as a course of study. The PI intends to use funds awarded to provide summer stipends for the S-STEM scholars to engage them in research projects as early as during the summer of their first year.