Kinetics and structural analysis of Ebony, a glial protein in the visual system of Drosophila

果蝇视觉系统中的胶质蛋白 Ebony 的动力学和结构分析

• 批准号: 146781222
• 负责人: Professor Dr. Bernhard T. Hovemann
• 金额: --
• 依托单位: Arbeitsgruppe Molekulare Zellbiochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/146781222?language=en
• 关键词: Kinetics; structural; analysis; Ebony; glial

In the visual system of arthropods histamine (HA) is the neurotransmitter for stimulus transmission from retina photoreceptor neurons to downstream postsynaptic neurons. We propose a recycling pathway for this neurotransmitter in Drosophila melanogaster (D. melanogaster) in which the two enzymes Ebony and Tan together with yet to be identified transporters hold key functions: HA from the synaptic cleft is fused to β-alanine by Ebony in the surrounding epithelial glia for inactivation. After transfer into photoreceptor neurons, β-alanyl-histamine or carcinine (CA) is hydrolyzed by Tan thereby replenishing the HA pool for vesicle loading. This working model strongly depends among others on the cata-lytic efficiency of Ebony. To verify the proposed recycling pathway with a key function of Ebony in neurotransmitter inactivation, it is essential to determine its reaction kinetics. Furthermore, enzyme structure will be solved to understand how specific recruitment of β-alanine and peptide bond formation can be performed in concert with fast HA transport into epithelial glia. As a prerequisite new expression systems and purification strategies will be employed that overcome the hitherto limited capability in preparing pure and active Ebony protein.

在节肢动物的视觉系统中，组胺（HA）是将刺激从视网膜感光神经元传递到下游突触后神经元的神经递质。我们提出了这种神经递质在果蝇（D。黑腹动物），其中两种酶Ebony和Tan与尚待鉴定的转运蛋白一起具有关键功能：来自突触间隙的HA通过Ebony与周围上皮神经胶质中的β-丙氨酸融合以失活。在转移到光感受器神经元中后，β-丙氨酰-组胺或癌碱（CA）被Tan水解，从而补充用于囊泡加载的HA库。这种工作模式在很大程度上取决于乌木的催化效率。为了验证所提出的回收途径与乌木在神经递质失活的关键功能，它是必不可少的，以确定其反应动力学。此外，酶结构将被解决，以了解β-丙氨酸的特异性募集和肽键形成如何与HA快速转运到上皮神经胶质中一起进行。作为一个先决条件，将采用新的表达系统和纯化策略，克服迄今有限的能力，在制备纯的和活性的乌木蛋白。

项目成果

Endogenous ciliary neurotrophic factor modulates anxiety and depressive-like behavior

• DOI: 10.1016/j.bbr.2012.01.020
• 发表时间: 2012-04
• 期刊: Behavioural Brain Research
• 影响因子: 2.7
• 作者: Isabella Peruga;S. Hartwig;D. Merkler;J. Thöne;B. Hovemann;G. Juckel;R. Gold;R. Linker