MicroCal Auto-iTC200; automated high sensitivity isothermal titration calorimetry

描述（由申请人提供）：获得MicroCal Auto-iTC系统将支持美国国立卫生研究院资助的项目，这些项目来自UAB校园和Emory校区的pi，需要对生物大分子的分子相互作用进行定量和完整的热力学评估。代表7个部门和4所学校的4个主要用户和8个次要用户以及代表3个部门和2所学校的7个正在等待NIH拨款或其他支持的次要用户将使用该仪器。等温滴定量热法（ITC）已经成功地应用于许多应用中，以研究各种系统，包括我们的用户组在这里描述的那些系统，例如，小分子，蛋白质，抗体，核酸和脂质的相互作用；突变和分子结构变化对结合机制的影响以及结合对结合结构的影响。ITC的独特之处在于它能够在单个实验中量化亲和、化学计量和结合的热力学参数(？H ?年代,?G在一个实验中；？Cp（3个实验）。没有其他技术不能在如此短的时间内提供如此广泛的信息，也没有如此少的材料来进行溶液中的相互作用，而且不需要外部标记。MicroCal Auto-iTC200系统是MicroCal提供的最新一代ITC，它在灵敏度和自动化方面无与伦比；事实上，没有其他商业上可用的机器人ITC，这一特性使其成为多用户核心设施的理想选择。首席研究员布鲁耶特博士和她的高级研究助理普罗塔塞维奇博士都有大约30年的量热学经验。他们目前管理着UAB综合癌症设施内的生物分子分析核心，该设施为用户提供Biacore2000和MicroCal自动capDSC。这个功能核心，目前的用户遍布校园，位于生物物理科学与工程中心（CBSE）内。如果目前的拨款获得资助，AutoiTC200系统也将通过这个现有的核心设施安置和管理。基础设施支持和空间由CBSE提供。将收取合理的用户费用，由经验丰富的人员进行日常维护和监督。额外援助将以额外费用提供。作为该仪器将支持的生物医学研究广度的一个例子，NIH为四个主要用户提供了资助标题：NIAID U01“炭疽、鼠疫和兔热病药物的发现和临床前开发”；变异链球菌Agl/II上SAG结合基序的测定NCI P50项目1“乳腺癌中的孢子”；NIGMS R01“线粒体蛋白易位的结构和功能研究”和NIDDK“Hsp40的结构和功能研究”。

DESCRIPTION (provided by applicant): Acquisition of the MicroCal Auto-iTC System will support NIH-funded projects from PIs across the UAB campus and Emory who require a quantitative and complete thermodynamic evaluation of the molecular interactions of biological macromolecules. Four major and 8 minor users who represent 7 departments and 4 schools and an additional 7 minor users with pending NIH grants or other support representing 3 departments and 2 schools will be served by this instrument. Isothermal titration calorimetry (ITC) has been successfully used in many applications to study a diverse range of systems including those proposed by our user group as described here, e.g., the interactions of small molecules, proteins, antibodies, nucleic acids and lipids; the effects of mutations and molecular structure changes on binding mechanism and the effect of binding on bound structure. ITC is unique in its ability to quantitate the affinity, stoichiometry and the thermodynamic parameters of binding in a single experiment (?H, ?S, ?G in one experiment; ?Cp in ~3 experiments). There simply is no other technique that can not provide the breadth of information in so little time nor with so little material for interactions in solution and furthermore no extrinsic labeling is required. The MicroCal Auto-iTC200 System is the newest generation ITC offered by MicroCal and it is unrivaled with respect to its sensitivity and automation; in fact, there is no other robotic ITC commercially available and this feature makes it ideal for a multi-user core facility. The PI, Dr. Brouillette, and her senior research associate, Dr. Protassevitch, each have about 30 years of calorimetry experience. They currently manage a biomolecular analysis core within UAB's Comprehensive Cancer Facility which offers a Biacore2000 and a MicroCal automated capDSC to users. This functioning core, with current users across campus, is physically housed within the Center for Biophysical Sciences and Engineering (CBSE). If the present grant is funded, the AutoiTC200 System will also be housed and administered through this existing core facility. Infrastructure support and space is provided by the CBSE. A reasonable user fee will be charged to cover routine maintenance and oversight by the experienced personnel. Additional assistance will be provided at an additional cost. As an example of the breadth of biomedical research which would be supported by this instrument, the NIH grant titles for the four major users are given: NIAID U01 "Discovery and Preclinical Development of Drugs for Anthrax, Plague and Tularemia;" NIDCR R01 "Determination of the SAG Binding Motif on Agl/II of Streptococcous Mutans;" NCI P50 project 1 "SPORE in Breast Cancer;" NIGMS R01 "Structural and Functional Studies for Mitochondrial Protein Translocations;" and NIDDK "Structural and Functional Studies of Hsp40."