A liquid chromatography system with integrated multi-angle light scattering detection to characterize protein folding, assembly, and modification

该应用程序请求资金购买与合适的尺寸排除层析系统(SEC-MALS)耦合的多角度光散射检测器。SEC-MALS提供了生物大分子的分子质量和回转半径的直接测量。它还允许在纯化过程中对分子进行表征，这对不稳定和对时间敏感的样品至关重要。这些能力在大西洋加拿大并不存在，迫切需要这些能力来提高申请者的研究能力。 所有五名共同申请者和他们目前的23名实习生，从本科生到博士后研究员，都将立即利用这一工具研究生命的基本过程和生物材料开发。SEC-MALS是一种确定蛋白质纯度和同质性的黄金标准技术，所有申请者都将常规使用该技术来监测其研究样本的质量。收购SEC-MALS仪器还将使研究丝素、丝状酶和真菌蛋白质自组装的新方向成为可能，这些蛋白质通常处于动态平衡状态，涉及几个低聚状态。其他研究项目将依靠SEC-MALS的优势，通过内含子剪接和泛素化有效地检测和量化蛋白质修饰，以及监测涉及转录因子的蛋白质-蛋白质相互作用。 该仪器将安装在Dalhousie的生物物理分析设施中，使所有共同申请者、他们的受训者以及哈利法克斯地区的其他研究人员能够不受限制地访问。该仪器将由学生和其他高素质人员直接操作，作为其研究培训的一部分，每个人都将在实际使用仪器和对所产生的数据进行专家分析方面得到指导。

This application requests funds to purchase a multi-angle light scattering detector coupled to a suitable size exclusion chromatography system (SEC-MALS). SEC-MALS provides a direct measurement of the molecular mass and radius of gyration of biomacromolecules. It also allows for molecules to be characterized during the purification process, which is essential for unstable and time-sensitive samples. These capabilities do not exist in Atlantic Canada, and are urgently needed to enhance the applicants' research capabilities. All five co-applicants and their 23 current trainees, ranging from undergraduate student to postdoctoral fellow, will immediately take advantage of this instrument for the study of fundamental processes of life and biomaterial development. SEC-MALS is a gold-standard technique to determine protein purity and homogeneity, and will be routinely used by all of the applicants to monitor the quality of their research samples. Acquisition of a SEC-MALS instrument will also enable new directions of study that investigate the self-assembly of silk, filamentous enzymes, and fungal proteins, where these proteins are often in a dynamic equilibrium involving several oligomeric states. Other research projects will rely upon the benefits of SEC-MALS to efficiently detect and quantify protein modification by intein splicing and ubiquitination, as well as monitoring protein-protein interactions involving transcription factors. This instrument will be installed in the Biophysical Analysis Facility at Dalhousie, providing unfettered access to all of the co-applicants, their trainees, and to other researchers in the Halifax region. This instrument will be operated directly by students and other highly qualified personnel as a part of their research training, each of whom will be mentored in hands-on use of the instrumentation and in expert analysis of the resulting data.