Targeting a Membrane Protease that Controls NK Cell Maturation

靶向控制 NK 细胞成熟的膜蛋白酶

• 批准号: 10631217
• 负责人: Joel L Pomerantz
• 金额: $ 20.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10631217
• 关键词: Alzheimer' s Disease; Antiviral Response; Aspartate; Aspartic Endopeptidases; Binding; Biological; Biological Assay; Cancerous; Cell Lineage; Cell Maturation; Cell physiology; Cells; Cellular Assay; Defect; Development; Disease; Effector Cell; Engineering; Event; Exhibits; Family; Health; Human; ITGAM gene; Immune; Immune system; In Vitro; Knock-in Mouse; Knowledge; Leukocytes; Lipid Bilayers; Lymphocyte; Lymphocyte Function; Lymphoid Cell; Malignant Neoplasms; Membrane; Metalloproteases; Molecular; Molecular Target; Mouse Strains; Mus; Natural Killer Cells; Pathway interactions; Peptide Hydrolases; Process; Protein Analysis; Proteins; Proteolysis; Role; Serine Protease; Site; Sorting; Stress; Substrate Interaction; T-Lymphocyte; Tumor Cell Line; arm; cell behavior; cytotoxic; cytotoxicity; design; immune system function; in vivo; mutant; neoplastic cell; novel; novel therapeutics; pathogen; presenilin; rhomboid; signal peptide peptidase; therapy design; tumor

PROJECT SUMMARY/ABSTRACT A variety of important biological pathways in health and disease are regulated by intramembrane proteolysis, the process that achieves the targeted cleavage of protein substrates within or proximal to the lipid bilayer. Four classes of intramembrane proteases have been described, including the site 2 protease (S2P) metalloprotease family, the rhomboid serine protease family, the Rce1 glutamyl protease family, and the aspartyl protease family that includes presenilins and the related Signal Peptide Peptidase (SPP)-like (SPPL) subfamily. The aspartyl proteases of the SPPL subfamily, including SPP, SPPL2a, SPPL2b, SPPL2c, and SPPL3, are perhaps the least understood subset of intramembrane proteases. Only recently have the biological roles of some of these proteases emerged, and only a handful of relevant biological substrates have been identified. How these intramembrane proteases specifically recognize and cleave their substrates remains mysterious. We discovered a critical role for SPPL3 in NK cell maturation and cytotoxicity. SPPL3 is required in a cell-autonomous manner for the maturation of NK cells from the immature CD27+CD11b- stage to the CD27+CD11b+ and CD27-CD22b+ stages, and for normal NK cell cytotoxicity toward tumor cell targets. Mice engineered to express only SPPL3 D271A in the NK lineage revealed that the proteolytic function of SPPL3 is required in NK cell maturation and function. Like other aspartyl intramembrane proteases of the SPPL subfamily, very little is known about how SPPL3 recognizes and cleaves its substrates. All SPPL proteases possess YD and GXGD motifs that contain the catalytic aspartates, but other regions in the protein that are required for substrate recognition and cleavage have not been identified. Furthermore, although some SPPL3 substrates have been identified, the relevant substrate that must be cleaved by SPPL3 during NK cell maturation is currently unknown. To expand our basic understanding of SPPL3 and related aspartyl intramembrane proteases, and to advance understanding of the key checkpoint in NK cell maturation controlled by SPPL3, we will use newly developed substrate binding and cleavage assays to define SPPL3 determinants required for substrate selection and to identify SPPL3 substrates in NK cells. Our results will expand understanding of intramembrane aspartyl proteases and illuminate molecular events that control NK cell development and function.

项目摘要/摘要 健康和疾病中各种重要的生物学途径受膜内蛋白水解的调节， 实现蛋白质底物的靶向裂解或与脂质双层的近端的过程。四个 已经描述了膜内蛋白酶的类别，包括位点2蛋白酶（S2P）金属蛋白酶 家族，菱形丝氨酸蛋白酶家族，RCE1谷氨酸蛋白酶家族和as植基蛋白酶家族 其中包括寄生虫和相关的信号肽肽酶（SPP） - 样（SPPL）亚家族。天冬氨酸 SPPL亚家族的蛋白酶，包括SPP，SPPL2A，SPPL2B，SPPL2C和SPPL3，也许是最少的 可理解膜内蛋白酶的子集。直到最近才有其中一些的生物学作用 出现蛋白酶，仅确定了少数相关的生物底物。这些如何 膜内蛋白酶专门识别并裂开其底物仍然神秘。我们发现了 SPPL3在NK细胞成熟和细胞毒性中的关键作用。 SPPL3是需要以细胞自治方式 为了使NK细胞从未成熟的CD27+CD11b-阶段到CD27+CD11b+和CD27-CD22B+的成熟 阶段，对于正常的NK细胞细胞毒性对肿瘤细胞靶标。设计仅表达SPPL3的小鼠 NK谱系中的D271a表明，在NK细胞成熟和 功能。像SPPL亚科的其他天冬氨酸膜内蛋白酶一样，对如何如何了解 SPPL3识别并切割其底物。所有SPPL蛋白酶都有YD和GXGD图案 催化天冬氨酸，但是蛋白质中底物识别和裂解所需的其他区域 尚未确定。此外，尽管已经确定了一些SPPL3底物，但相关 目前尚不清楚必须在NK细胞成熟过程中被SPPL3切割的底物。扩大我们的基本 了解SPPL3和相关的天冬氨酸膜内蛋白酶，并促进对 由SPPL3控制的NK细胞成熟中的关键检查点，我们将使用新开发的底物绑定和 切割分析要定义底物选择所需的SPPL3决定因素并识别SPPL3底物 在NK细胞中。我们的结果将扩大对膜内天冬氨酸蛋白酶和照明分子的了解 控制NK细胞开发和功能的事件。