Chemical Biology of Transglutaminases

转谷氨酰胺酶的化学生物学

• 批准号: RGPIN-2019-05893
• 负责人: Keillor, Jeffrey
• 金额: $ 5.76万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748193
• 关键词: Chemical; Biology; Transglutaminases

Enzymes are proteins that catalyse almost all of the chemical reactions necessary to support life. Enzymes are known to show a high degree of specificity with respect to the molecules that they recognize and transform, known as substrates. In order to understand the biological role of an enzyme, it is essential to first understand its specificity, and the nature of the transformation it catalyses. The transglutaminases (TGases) are a family of eight enzymes that have been implicated in a wide variety of important physiological processes. However, we still have a limited understanding of which of these different TGases catalyse which reactions, and with which substrates. For example, they all catalyse either the cross-linking of different proteins, or the reversible attachment of biological amines to those proteins, but we don't know which TGases modify which proteins, in what way, and in which tissues. The Keillor group is uniquely qualified to answer these questions. We have many years' experience studying tissue TGase (TG2), and have recently discovered how to make small molecules that will react specifically with TG2, and not with other TGases. This has led to a breakthrough in understanding how TG2 plays a role in cancer metastasis. Over the next granting period, we will apply the same approach, to make molecules that will react specifically with three additional members of the TGase family: Factor XIII, TG1 and TG3. TG1 and TG3 are both thought to play roles in the stabilization of skin; however, the excess activity of one and the deficiency of the other have both been implicated in skin disorders. Factor XIII circulates in the blood and assists in the coagulation process, but it is also involved in bone development and in clotting disorders. For each of these four TGases, we will 1) determine their specificity, 2) develop novel methods for measuring their specific activity, 3) develop inhibitors for blocking their specific activity, and 4) develop molecular probes that will reveal where the enzymes and their substrates are located, in living cells. Achieving these objectives will allow us to dissect the related activities of these important enzymes, to discover their unique biological roles and to guide the future treatment of physiological disorders related to their unregulated activity. The detailed methods required to achieve these multidisciplinary objectives are numerous, but they are all inherent in our current research program. We routinely employ organic synthesis, molecular biology, protein biochemistry, bioanalytical techniques, kinetic studies, mammalian cell culture and fluorescence microscopy. Trainees in the Keillor group will therefore have the opportunity to be trained across a broad array of highly relevant techniques, rather than specializing in just one. This will prepare them very well for impactful positions in the modern biotech or pharmaceutical industries, or in academia.

酶是催化几乎所有维持生命所必需的化学反应的蛋白质。已知酶对它们识别和转化的分子（称为底物）显示出高度的特异性。为了了解酶的生物学作用，首先必须了解其特异性及其催化的转化性质。转谷氨酰胺酶（TGases）是一个由八种酶组成的家族，它们参与了多种重要的生理过程。然而，我们仍然有一个有限的了解，这些不同的TGases催化哪些反应，并与哪些底物。例如，它们都催化不同蛋白质的交联，或生物胺与这些蛋白质的可逆连接，但我们不知道哪些TGases修饰哪些蛋白质，以何种方式以及在哪些组织中。Keillor集团是唯一有资格回答这些问题的。我们有多年研究组织TGase（TG 2）的经验，最近发现了如何制造与TG 2特异性反应的小分子，而不是与其他TGases。这导致了对TG 2如何在癌症转移中发挥作用的理解的突破。在下一个授权期内，我们将采用相同的方法，制造与TGase家族的另外三个成员特异性反应的分子：因子XIII，TG 1和TG 3。 TG 1和TG 3都被认为在稳定皮肤中发挥作用;然而，一个的过度活性和另一个的缺乏都与皮肤疾病有关。因子XIII在血液中循环并协助凝血过程，但它也参与骨骼发育和凝血障碍。对于这四种TGases中的每一种，我们将1）确定它们的特异性，2）开发测量它们的比活性的新方法，3）开发用于阻断它们的比活性的抑制剂，以及4）开发将揭示酶及其底物在活细胞中的位置的分子探针。实现这些目标将使我们能够剖析这些重要酶的相关活动，发现它们独特的生物学作用，并指导未来与其不受调节的活动相关的生理疾病的治疗。实现这些多学科目标所需的详细方法有很多，但它们都是我们目前研究计划所固有的。我们通常采用有机合成，分子生物学，蛋白质生物化学，生物分析技术，动力学研究，哺乳动物细胞培养和荧光显微镜。因此，Keillor集团的学员将有机会接受广泛的高度相关技术培训，而不是只专注于一种技术。这将为他们在现代生物技术或制药行业或学术界的有影响力的职位做好准备。