NCL with induced a-Helices: de Novo Chemical Synthesis of the ATAD2 bromodomain

具有诱导α-螺旋的 NCL：ATAD2 溴结构域的从头化学合成

• 批准号: 8309682
• 负责人: Gardner Silas Creech
• 金额: $ 4.71万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8309682
• 关键词: Adoption; Amino Acid Motifs; Bromodomain; Chemicals; Coupling; Genetic Transcription; Glycopeptides; Goals; Growth; Human; Investigation; Length; Ligation; Lung Neoplasms; Malignant Neoplasms; Mammary Neoplasms; Mitosis; Molecular Conformation; Patients; Peptide Fragments; Peptide Synthesis; Peptides; Phase; Prostatic Neoplasms; Protein Fragment; Proteins; Protocols documentation; Reaction; Secondary Protein Structure; Solid; Solutions; Structure; Technology; Testing; Up-Regulation; Vertebral column; alpha helix; base; chemical synthesis; design; outcome forecast; ovarian neoplasm; peptide chemical synthesis; polypeptide; prevent; protein aminoacid sequence; synthetic protein; therapeutic target; tumor; tumor growth

DESCRIPTION (provided by applicant): The de novo chemical synthesis of proteins has the potential to rapidly accelerate the study of proteins by providing rapid access to natural and rationally designed unnatural proteins. Currently, however, chemical synthesis of proteins is limited to relatively small sized polypeptides and glycopeptides, due to a number of practical factors. Generally, the efficiency of ligation of peptide fragments decreases with increasing peptide length, where peptide aggregation is an often observed obstacle. We propose an alternative paradigm for the protection of peptides against aggregation and the size limitations of native ligation, in contrast to the introduction of extraneous protection functionalities and auxiliaries. The induction of secondary structure in solution for peptide fragments that are predisposed to the formation of alpha-helices should serve to protect the peptides against destructive aggregation by rigidifying the peptide backbone into a compact conformation. This line of reasoning is in stark contrast to the typical native chemical ligation protocol that is historically performed exclusively under denaturing conditions. In addition to reduced propensity towards aggregation, we expect the helical protein fragments to have increased efficiency of ligation due to the conformation compactness and rigidity, and a corresponding reduction in the negative correlation between peptide complexity and ligation efficiency. In order to investigate this approach towards peptide ligation, the bromodomain of human protein ATAD2 will be synthesized. In only the past few years, this protein has been identified as upregulated in breast, prostate, lung, and ovarian tumors, and numerous studies have correlated the over expression of ATAD2 with cancer growth and patient prognosis. Considering the involvement of ATAD2 with protein transcription and mitosis along with the upregulation observed in a wide variety of tumor types, it is no surprise that ATAD2 has been identified as a potential therapeutic target. The bromodomain of ATAD2 is an ideal target to test the helical stabilization hypothesis because it is made up of five alpha-helices, each of ideal size for solid phase peptide synthesis, and includes two relatively hydrophobic regions with potential for aggregation. This investigation has the potential for broad implications, as the strategic revision for the chemical synthesis of proteins that is proposed applies new rules for the disconnection of polypeptides into their corresponding fragments.

描述（由申请人提供）：蛋白质的从头化学合成具有通过提供快速获得天然和合理设计的非天然蛋白质来快速加速蛋白质研究的潜力。然而，目前，由于许多实际因素，蛋白质的化学合成限于相对小尺寸的多肽和糖肽。通常，肽片段的连接效率随着肽长度的增加而降低，其中肽聚集是经常观察到的障碍。我们提出了一种替代的范式，保护肽对聚集和大小限制的天然连接，在引入外来的保护功能和助剂。对于易于形成α-螺旋的肽片段，在溶液中诱导二级结构应通过将肽骨架刚性化成紧凑构象来保护肽免于破坏性聚集。这一推理路线与历史上仅在变性条件下进行的典型天然化学连接方案形成鲜明对比。除了降低的聚集倾向之外，我们预期螺旋蛋白片段由于构象紧密性和刚性而具有增加的连接效率，以及肽复杂性和连接效率之间的负相关性的相应降低。为了研究这种肽连接方法，将合成人蛋白ATAD 2的溴结构域。仅在过去的几年中，这种蛋白质已被确定为在乳腺、前列腺、肺和卵巢肿瘤中上调，并且许多研究将ATAD 2的过度表达与癌症生长和患者预后相关联。考虑到ATAD 2参与蛋白质转录和有丝分裂，沿着在多种肿瘤类型中观察到的上调，ATAD 2已被鉴定为潜在的治疗药物就不足为奇了 目标ATAD 2的布罗莫结构域是测试螺旋稳定化假设的理想靶标，因为它由五个α-螺旋组成，每个α-螺旋具有用于固相肽合成的理想尺寸，并且包括两个具有聚集潜力的相对疏水区域。这项调查具有广泛的影响的潜力，作为提出的蛋白质的化学合成的战略性修订适用于新的规则断开多肽成其相应的片段。