Biophysical and Model Studies of NiSOD

NiSOD 的生物物理和模型研究

• 批准号: 0321482
• 负责人: Michael Maroney
• 金额: $ 45.4万
• 依托单位: University of Massachusetts Amherst
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0321482&HistoricalAwards=false
• 关键词: Biophysical; Model; Studies; NiSOD

Nickel-containing superoxide dismutases (NiSODs) represent a completely novel mechanism for the elimination of superoxide in biological systems. The proposed project uses an interdisciplinary approach to explore the structure and function of NiSODs found in Streptomyces species, an organism that is responsible for many of the antibiotics currently available to medicine. Single crystal x-ray diffraction, x-ray absorption spectroscopy, vibrational spectroscopy and mass spectrometry will be used to examine structural aspects of NiSOD. These techniques will characterize the overall structure of the protein, determine the structure of the metal site and the dioxygen species present in various forms of the enzyme (including potential reaction intermediates or inhibitor complexes), and help to elucidate the role of quaternary structure in the function of the enzyme. Site-directed mutagenesis techniques and kinetic measurements will be used to test the role of specific amino acid residues in the structure of the catalytic site, with the effect of mutation monitored by measurement of the catalytic rate constant. Kinetic methods will also be used to examine the role of quaternary structure and an unusual product inhibition that is associated with peroxide in NiSOD.Broader Impact: The proposed research also provides a significant training opportunity for students at all levels (undergraduate through post-doctoral) at the interface of biochemistry and inorganic chemistry. Students will benefit from the association with a Chemistry-Biology Interface training program on campus, collaborations with scientists at other institutions in the U.S, and from a NSF-funded exchange program with a laboratory in Hungary. Students will be exposed to a number of techniques and concepts from both biochemistry and inorganic chemistry. Students will be drawn from a diverse pool, and effort will be made to provide opportunities for minorities.This project is funded jointly by Molecular Biochemistry and Inorganic, Bioinorganic & Organometallic Chemistry Program.

含镍超氧化物歧化酶（NiSODs）是一种消除生物系统中超氧化物的全新机制。拟议的项目使用跨学科的方法来探索链霉菌物种中发现的nisod的结构和功能，链霉菌是目前医学上可用的许多抗生素的微生物。单晶x射线衍射、x射线吸收光谱、振动光谱和质谱将用于检查NiSOD的结构方面。这些技术将表征蛋白质的整体结构，确定金属位点的结构和存在于各种形式的酶（包括潜在的反应中间体或抑制剂复合物）中的双氧物种，并有助于阐明四级结构在酶的功能中的作用。位点定向诱变技术和动力学测量将用于测试特定氨基酸残基在催化位点结构中的作用，通过测量催化速率常数来监测突变的效果。动力学方法也将用于研究在NiSOD中与过氧化物相关的季元结构和不寻常的产物抑制的作用。更广泛的影响：拟议的研究还为生物化学和无机化学界面的各级学生（本科生到博士后）提供了重要的培训机会。学生将受益于与校园化学-生物学界面培训项目的联系，与美国其他机构的科学家合作，以及与匈牙利实验室的nsf资助的交流项目。学生将接触到生物化学和无机化学的一些技术和概念。学生将从多样化的人群中选拔，并将努力为少数民族提供机会。本项目由分子生物化学与无机、生物无机及有机金属化学专业联合资助。