QPP: Protease that Prevents Apoptosis in Quiescent Cells

QPP：防止静止细胞凋亡的蛋白酶

• 批准号: 7162147
• 负责人: Brigitte T. Huber
• 金额: $ 30.06万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7162147
• 关键词: Activated Lymphocyte; Active Sites; Affinity; Apoptosis; Biological Assay; Cell Cycle; Cell Death; Cells; Chromatography; Cleaved cell; Collaborations; Dipeptidases; Dipeptides; Dominant-Negative Mutation; Embryo; Endopeptidases; Enhancers; Enzymes; Eukaryotic Cell; Gene Chips; Gene Silencing; Genes; Genetic Transcription; Genome; Genomics; Goals; Half-Life; Human; Immune system; In Situ; In Vitro; Individual; Induction of Apoptosis; Interphase Cell; Introns; Lead; Libraries; Link; Lymphocyte; Maps; Mediating; Monitor; Mus; Mutant Strains Mice; N-terminal; Neurons; Organ; Peptide Hydrolases; Peptides; Phenotype; Physiological; Planet Mars; Process; Proline; Proteins; Regulation; Reporter; Rest; Reverse Transcriptase Polymerase Chain Reaction; Role; STAT5A gene; Sequence Analysis; Site; Specificity; Structure; System; TP53 gene; Tamoxifen; Testing; Time; Tissues; Transfection; Transgenic Mice; Up-Regulation; c-myc Genes; cell type; combinatorial; embryonic stem cell; enzyme activity; gel electrophoresis; homologous recombination; in vivo; membrane dipeptidase; mutant; novel; prevent; programs; promoter; receptor; recombinase; research study; transcription factor; transmission process

QPP (Quiescent cell _.Proline di-_Peptidase), also termed DPP2/7, is a vesicular protease that cleaves di- peptides from the N-terminus of proteins, thereby impacting their half-life and/or receptor specificity. While the full functional profile of this enzyme is far from understood, in vitro experiments demonstrate that QPP is a major regulator of the Go quiescence program in primary lymphocytes and neuronal cells. The primary objective of this renewal application is to analyze the mechanism of QPP-mediated survival in resting lymphocytes. Aim I: The most direct approach to define the functional role of QPP is to generate a mutant mouse deficient in QPP. Since previous attempts to derive conventional QPP# mice were curbed by embryonic lethality, we propose a novel alternative to generate the mutant mouse, utilizing the Cre/IoxP recombinase system that allows conditional and inducible genome alterations. Aim I1" Inhibition of QPP advances quiescent lymphocytes into cell cycle, leading to upregulation of c- Myc and p53 and finally apoptosis induction. Thus, it is postulated that QPP enzyme activity is required for maintaining the Go program. To test this hypothesis, the control of expression and mechanism of action of QPP will be delineated. Special emphasis will be given to LKLF and STAT5, two transcription factors that are implicated in positive and negative regulation of QPP expression, respectively. Furthermore, the role of c-Myc and p53 in the apoptosis pathway will be analyzed. Aim II1: To fully understand the Go survival program in eukaryotic cells, it is essential to define the physiological substrate(s) of QPP. Since this is the most ambitious project, various approaches will be used to reach this goal, including the screen of combinatorial dipeptide substrate libraries, the structural comparison of human and murine QPP with that of DPP4, and the isolation of a physiological substrate(s) by affinity matrix chromatography. Collectively, these studies will augment our limited understanding of the constitutive Go survival program in eukaryotic cells.

QPP（Quiescent cell _Proline di_Peptidase），也称为DPP 2/7，是一种切割二-脯氨酸的囊泡蛋白酶。 从蛋白质的N-末端的肽，从而影响它们的半衰期和/或受体特异性。 虽然这种酶的完整功能概况还远未了解，但体外实验表明， QPP是初级淋巴细胞和神经元细胞中Go静止程序的主要调节剂。 本次更新申请的主要目的是分析QPP介导的存活机制 在静止的淋巴细胞中。目的一：界定品质专业计划职能的最直接方法是 产生QPP缺陷的突变小鼠。由于先前尝试获得常规QPP#小鼠 受到胚胎致死性的抑制，我们提出了一种新的替代方法来产生突变小鼠， 利用允许条件性和诱导性基因组改变的Cre/IoxP重组酶系统。 抑制QPP可使静止淋巴细胞进入细胞周期，导致c- Myc和p53以及最后的凋亡诱导。因此，假定需要QPP酶活性 来维护围棋程序为了验证这一假设，我们研究了对基因表达的控制和基因表达的机制。 将描述QPP的作用。特别强调将给予LKLF和STAT 5，两个转录 分别涉及QPP表达的正性和负性调节的因子。 此外，c-Myc和p53在凋亡途径中的作用将被分析。目标二1：充分 为了了解真核细胞中的Go生存程序，定义生理学上的 QPP的底物。由于这是最雄心勃勃的项目，将采用各种方法来实现 这一目标，包括筛选组合二肽底物库， 人和鼠QPP与DPP 4的同源性，以及通过亲和层析分离生理底物 基质色谱法总的来说，这些研究将增加我们对 真核细胞中的组成型Go生存程序。

项目成果

Th17 differentiation is the default program for DPP2-deficient T-cell differentiation.

• DOI: 10.1002/eji.201041157
• 发表时间: 2011-06
• 期刊: EUROPEAN JOURNAL OF IMMUNOLOGY
• 影响因子: 5.4
• 作者: Mele, Deanna A.;Sampson, James F.;Huber, Brigitte T.
• 通讯作者: Huber, Brigitte T.

Biologically distinct conformations of Bcl-x can be resolved using 2D isoelectric focusing.

Bcl-x 的生物学独特构象可以使用 2D 等电聚焦来解析。

• DOI: 10.1016/j.molimm.2009.02.031
• 发表时间: 2009
• 期刊: Molecular immunology
• 影响因子: 3.6
• 作者: Rockwell,KarenR;Huber,BrigitteT
• 通讯作者: Huber,BrigitteT

Aminodipeptidase inhibitor-induced cell death in quiescent lymphocytes: a review.

氨基二肽酶抑制剂诱导的静止淋巴细胞细胞死亡：综述。

• DOI: 10.1023/a:1009675223443
• 发表时间: 2000
• 期刊: Apoptosis : an international journal on programmed cell death.
• 作者: Chiravuri,M;Huber,BT
• 通讯作者: Huber,BT