Determinants of Rod and Cone Response Characteristics

杆和锥响应特性的决定因素

• 批准号: 7455001
• 负责人: JAMES Bryant HURLEY
• 金额: $ 36.69万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7455001
• 关键词: Address; Apoptosis; Biochemical Reaction; Calcium; Characteristics; Condition; Consensus Sequence; Dark Adaptation; Data; Funding; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GRK1 gene; Goals; Kinetics; Light; Lighting; Mass Spectrum Analysis; Methodology; Methods; Mus; Night Blindness; Phosphorylation; Phosphorylation Site; Photoreceptors; Physiological; Protein Dephosphorylation; Reaction; Retina; Retinal Cone; Retinal Pigments; Rhodopsin; Signal Pathway; Site; Testing; Vertebrate Photoreceptors; Vision; Visual; Zebrafish; base; improved; preference; receptor; recoverin protein; response; rhodopsin kinase; visual cycle

DESCRIPTION (provided by applicant): Phosphorylation of visual pigments is essential for normal vision. Deficiencies in this biochemical reaction can cause night blindness and photoreceptor apoptosis. Phosphorylation uncouples rhodopsin and most G-protein coupled receptors from downstream signaling pathways. The sites of phosphorylation on most G-protein coupled receptors are not known. During the previous funding period we developed a mass- spectrometry based method that enables us to determine the sites and kinetics of phosphorylation and dephosphorylation reactions that occur on visual pigments in intact retinas. We have used the method to establish the sites and kinetics of phosphorylation on mouse and zebrafish visual pigments under a variety of physiological conditions. We propose to use the method to pursue four new specific aims. 1. Evaluate rhodopsin phosphorylation during long-term illumination and identify relationships between visual pigment phosphorylation, visual cycle and visual sensitivity. 2. Evaluate factors that influence phosphorylation of cone visual pigments. 3. Identify structural characteristics that determine site selection and kinetics of phosphorylation by rhodopsin kinase. 4. Evaluate the influence of recoverin and Calcium on visual pigment phosphorylation. These studies will provide the most precise data yet available about the kinetics and site preferences for phosphorylation of G-protein coupled receptors.

描述（由申请人提供）：视色素的磷酸化对正常视力至关重要。这种生化反应的缺陷会导致夜盲症和感光细胞凋亡。磷酸化使视紫红质和大多数G蛋白偶联受体与下游信号通路解偶联。大多数G蛋白偶联受体的磷酸化位点尚不清楚。在之前的资助期间，我们开发了一种基于质谱的方法，使我们能够确定完整视网膜中视色素上发生的磷酸化和去磷酸化反应的位点和动力学。我们已经使用的方法，以建立在各种生理条件下的小鼠和斑马鱼的视色素磷酸化的网站和动力学。 我们建议使用该方法来追求四个新的具体目标。 1.评估长期光照下视紫红质磷酸化，并确定视色素磷酸化，视觉周期和视觉敏感度之间的关系。 2.评估影响视锥细胞色素磷酸化的因素。 3.识别决定视紫红质激酶磷酸化位点选择和动力学的结构特征。 4.评价恢复素和钙对视觉色素磷酸化的影响。 这些研究将提供最精确的数据，尚未获得的动力学和位点偏好的磷酸化G蛋白偶联受体。