Molecular screens for pVHL-associated isopeptidase VDU1

pVHL 相关肽酶 VDU1 的分子筛选

• 批准号: 6941091
• 负责人: Michael R Mattern
• 金额: $ 24.31万
• 依托单位: PROGENRA, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6941091
• 关键词: Von Hippel Lindau syndrome; antineoplastics; biotechnology; drug discovery /isolation; drug screening /evaluation; endopeptidases; enzyme activity; enzyme inhibitors; high throughput technology; technology /technique development; ubiquitin

DESCRIPTION (provided by applicant): VDU1 is a recently described ubiquitin isopeptidase that binds to and is ubiquitinated by the E3 ligase complex of the vonHippel-Lindau protein (pVHL). VDU1 has been found to be a target of ubiquitination (and proteasomal degradation) by the tumor suppressor protein complex VCB-CUL2 (an E3 ubiquitin ligase complex consisting of the von Hippel-Lindau protein pVHL, elongin C, elongin B, and cullin-2 (Li, Na et al. 2002; Li, Wang et al. 2002). pVHL is mutated in certain cancers and behaves as a tumor suppressor gene. Biochemical and genetic evidence suggests that VDU1, along with a limited number of other ubiquitination targets of the pVHL E3 ligase, has a role in establishing or maintaining the transformed state of pVHLmutant tumors. One of the E3 ligase's identified target proteins, HIFa, is known to induce angiogenic factors under certain conditions, and thus acts as an oncogene product. The beta-domain region of pVHL, which is the site of naturally occurring mutations, is the locus of VDU1 interaction, and VDU1 can be coimmunoprecipitated in the VCB-CUL2 complex. Moreover, the ubiquitination and degradation of VDU1 by a pVHL-dependent pathway is abrogated by VHL mutations that disrupt interactions with VDU1. Thus, targeted degradation of VDU1 by pVHL may be important in suppressing tumor formation and/or maintenance, and VDU1 may have oncogenic activity that is uncovered in the absence of the functional ligase. In this proposed one-year Phase I, we will configure a novel isopeptidase assay for VDU1 and configure it for high throughput screening, with the aim using this assay to discover novel antitumor drugs that act by inhibiting the cellular activity that would normally be prevented by wild type pVHL. We will also screen VDU1 against other peptide substrates to assess its cleavage patterns. The ultimate commercial goal of the development of an assay for VDU1 is to discover a drug with efficacy as a single agent or a component of conjoint therapy against renal and other cancers. An assay for the related and highly homologous isopeptidase VDU2 will be developed in Phase II and hits from the VDU1 assay screened with this assay as well.

描述（由申请人提供）： VDU 1是最近描述的泛素异肽酶，其结合vonHippel-Lindau蛋白（pVHL）的E3连接酶复合物并被其泛素化。已经发现VDU 1是肿瘤抑制蛋白复合物VCB-CUL 2（由von Hippel-Lindau蛋白pVHL、延伸蛋白C、延伸蛋白B和cullin-2组成的E3泛素连接酶复合物）的泛素化（和蛋白酶体降解）的靶标（Li，Na等2002; Li，Wang等2002）。pVHL在某些癌症中突变，并表现为肿瘤抑制基因。生物化学和遗传学证据表明VDU 1，沿着pVHL E3连接酶的有限数量的其它泛素化靶点，在建立或维持pVHL突变肿瘤的转化状态中具有作用。已知E3连接酶的鉴定的靶蛋白之一HIF a在某些条件下诱导血管生成因子，并因此充当癌基因产物。pVHL的β-结构域区域是天然存在的突变位点，是VDU 1相互作用的位点，并且VDU 1可以在VCB-CUL 2复合物中共免疫沉淀。此外，通过PVHL依赖性途径的VDU 1的泛素化和降解被破坏与VDU 1的相互作用的VHL突变废除。因此，PVHL对VDU 1的靶向降解在抑制肿瘤形成和/或维持中可能是重要的，并且VDU 1可能具有在不存在功能性连接酶的情况下未被发现的致癌活性。在这项拟议的为期一年的第一阶段中，我们将配置一种新型的VDU 1异肽酶测定法，并将其配置用于高通量筛选，目的是使用该测定法发现新型抗肿瘤药物，这些药物通过抑制通常会被野生型阻止的细胞活性来发挥作用pVHL。我们还将针对其他肽底物筛选VDU 1以评估其切割模式。开发VDU 1测定法的最终商业目标是发现一种药物，该药物作为单一药物或联合治疗的组分对肾癌和其他癌症有效。将在II期开发相关和高度同源的异肽酶VDU 2的检测试剂盒，并使用该检测试剂盒筛选VDU 1检测试剂盒的命中结果。

项目成果

Deubiquitinating enzymes as novel anticancer targets.

• DOI: 10.2217/14796694.3.2.191
• 发表时间: 2007-04-01
• 期刊: Future oncology (London, England)
• 作者: Nicholson, Benjamin;Marblestone, Jeffrey G;Mattern, Michael R
• 通讯作者: Mattern, Michael R

Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities.

• DOI: 10.1110/ps.083450408
• 发表时间: 2008-06
• 期刊: Protein science : a publication of the Protein Society
• 作者: Nicholson B;Leach CA;Goldenberg SJ;Francis DM;Kodrasov MP;Tian X;Shanks J;Sterner DE;Bernal A;Mattern MR;Wilkinson KD;Butt TR
• 通讯作者: Butt TR