CYTOSOL/SECRETORY GRANULE LUMEN COMMUNICATION

细胞溶胶/分泌颗粒腔通讯

• 批准号: 6446870
• 负责人: LESLIE C BELL-PARIKH
• 金额: $ 2.41万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6446870
• 关键词: adrenocorticotropic hormone; amidation /deamidation; chimeric proteins; cytoplasm; cytoskeleton; enzyme activity; exocytosis; granule; membrane proteins; oxygenases; phosphoproteins; phosphorylation; pituitary gland; protein tyrosine kinase; recombinant proteins; tissue /cell culture

In many endocrine cells and most neurons, bioactive peptides are stored in large dense core vesicles (LDCVs) until release is triggered by an appropriate stimulus. Peptidylglycine alpha-amidating monooxygenase (PAM), a peptide biosynthetic enzyme, is found in nearly all cells with LDCVs and is one of the few peptide processing enzymes that spans the granule membrane. The overall goal of this research proposal is to investigate the role of Peptidylglycine alpha-Amidating Monooxygenase (PAM), a peptide biosynthetic enzyme, in signaling the cytoskeletal rearrangement required for secretion of peptides from large dense core vesicles (LDCVs), and to probe the potential for PAM to act as a transmembrane receptor, communicating the status of the LDCV lumenal milieu to relevant cytosolic factors. This will be investigated using a streptolysin-O permeabilized cell model to identify second messengers and/or PAM interactor proteins which stimulate this signaling event within the cytoplasm. The involvement of Kalirin, a recently identified PAM cytosolic domain interactor protein with GDP/GTP exchange factor activity, will be characterized using this permeabilized cell system. Further, chimeric PAM proteins will be used to screen granule components for ligands that may potentiate cytoskeletal rearrangement from within the granule lumen through a specific interaction with PAM.

在许多内分泌细胞和大多数神经元中，生物活性肽都存储在大型密集的核囊泡（LDCV）中，直到释放是由适当的刺激触发的。 在几乎所有具有LDCVS的细胞中发现了肽基甘氨酸αα酶（PAM），一种肽生物合成酶，是跨颗粒状膜的少数肽加工酶之一。 The overall goal of this research proposal is to investigate the role of Peptidylglycine alpha-Amidating Monooxygenase (PAM), a peptide biosynthetic enzyme, in signaling the cytoskeletal rearrangement required for secretion of peptides from large dense core vesicles (LDCVs), and to probe the potential for PAM to act as a transmembrane receptor, communicating the status LDCV Lumenal Milieu与相关的胞质因子。 这将使用链霉菌素-O通透性细胞模型进行研究，以鉴定第二个使者和/或PAM相互作用的蛋白质，从而刺激细胞质中的这种信号传导事件。 使用此通透性细胞系统将表征Kalirin的参与，Kalirin是最近鉴定出的PAM胞质结构域相互作用蛋白与GDP/GTP交换因子活性的参与。此外，嵌合PAM蛋白将用于筛选颗粒成分的配体，这些配体可能会通过与PAM的特定相互作用从颗粒腔内增强细胞骨架重排。