CAREER: Target Specificity of Cysteine Desulfurases in Bacillus Subtilis

职业：枯草芽孢杆菌中半胱氨酸脱硫酶的靶标特异性

• 批准号: 1054623
• 负责人: Patricia Dos Santos
• 金额: $ 76.49万
• 依托单位: Wake Forest University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1054623&HistoricalAwards=false
• 关键词: CAREER; Target; Specificity; Cysteine; Desulfurases

Intellectual MeritSulfur-containing cofactors participate in a wide range of essential biological reactions. Although sulfur has long been recognized as indispensable to protein cofactors, its incorporation into biomolecules is not completely understood. In most organisms, the recruitment of sulfur for the synthesis of several cofactors involves the participation of a single general enzyme, cysteine desulfurase. The recruitment of such a promiscuous enzyme therefore poses an experimental challenge for dissecting the specific mechanisms of sulfur mobilization because its inactivation affects the synthesis of most sulfur cofactors. Gram-positive microbes such as Bacillus subtilis, on the other hand, code for several distinct cysteine desulfurases. Based on their genomic location, coding sequences, and preliminary analyses, the lab hypothesizes that each has specialized functions in sulfur metabolism. Using a combination of molecular genetics, biochemical and spectroscopic approaches, the lab will investigate the specific catalytic mechanisms by which these enzymes mobilize sulfur and their participation in the biosynthesis of sulfur cofactors using Bacillus subtilis as the model organism. This project will (i) establish the evolutionary importance of the differentiation and specialization of cysteine desulfurases; (ii) define key aspects of their structural, mechanistic, and functional relationships; and (iii) provide an excellent model system to study sulfur-trafficking reactions uncoupled from iron-sulfur metabolism. Successful results will transform our understanding of life-sustaining reactions and the intersections of metabolic pathways in the synthesis of essential sulfur cofactors.Broader ImpactThe broader impacts of this CAREER project will include the involvement of graduate students from Wake Forest University and undergraduate students from neighboring institutions in a mentored research/professional development program. The support team comprised of graduate students and instructors will develop a career plan for the students, which will include summer research experience and participation in professional development activities and meetings during the academic year. This program aims to broaden the scientific research preparation and participation of underrepresented groups in science. The team-mentoring approach will also have an impact on participating graduate students and their training as future teacher-scholars.

智力优点含硫辅助因子参与广泛的重要生物反应。 尽管硫长期以来一直被认为是蛋白质辅助因子不可或缺的成分，但其​​与生物分子的结合尚不完全清楚。 在大多数生物体中，为合成几种辅因子而募集硫涉及单一通用酶——半胱氨酸脱硫酶的参与。 因此，这种混杂酶的招募对剖析硫动员的具体机制提出了实验挑战，因为它的失活会影响大多数硫辅因子的合成。另一方面，革兰氏阳性微生物（例如枯草芽孢杆菌）编码几种不同的半胱氨酸脱硫酶。根据它们的基因组位置、编码序列和初步分析，实验室假设每一种在硫代谢中都具有专门的功能。 该实验室将结合分子遗传学、生化和光谱方法，以枯草芽孢杆菌为模型生物，研究这些酶动员硫的具体催化机制及其参与硫辅助因子的生物合成。 该项目将 (i) 确定半胱氨酸脱硫酶的分化和特化的进化重要性； (ii) 定义其结构、机械和功能关系的关键方面； (iii)提供一个优秀的模型系统来研究与铁硫代谢无关的硫运输反应。 成功的结果将改变我们对生命维持反应以及必需硫辅因子合成中代谢途径交叉点的理解。 更广泛的影响 该职业项目的更广泛影响将包括维克森林大学的研究生和邻近机构的本科生参与指导研究/专业发展计划。 由研究生和教师组成的支持团队将为学生制定职业计划，其中包括暑期研究经验以及学年期间参加专业发展活动和会议。 该计划旨在扩大科学研究准备和科学领域代表性不足群体的参与。 团队指导方法还将对参与的研究生及其作为未来教师学者的培训产生影响。