LISST

这项研究计划以理解结构基础为目标 用于糖酵解酶共定位和靶向Z-Disk和M-Disk 肌节的线条作为一种理解这一本质的手段 糖酵解酶组织为肌肉提供能量 收缩。在果蝇的飞行肌肉中，糖酵解 甘油-3-磷酸脱氢酶(GPDH)，甘油醛-3- 磷酸脱氢酶(GAPDH)和醛缩酶定位于Z-DISS/M-。 台词。GAPDH和醛缩酶的定位依赖于同时 GPDH的本地化。在没有糖酵解酶的情况下- 在飞行肌肉中定位，苍蝇不能飞。进一步阐明 支持协同定位的蛋白质-蛋白质相互作用的本质 将有助于更好地理解ATP 糖酵解产生的产物支持肌肉功能。它还有待于 确定糖酵解酶是否物理上相互作用或是否 其他蛋白质在共定位中也发挥了作用。这一点将被确定 采用酵母双杂交系统和免疫共沉淀技术。 肌肉特异性异构体中发现的C末端三肽Q-N，L， GPDH目标需要GPDH-1。糖酵解的其他结构域 酶可能在将这些酶靶向Z-Disks/M- 线路，并将被识别。编码相互作用的蛋白质的基因 与GPDH和醛缩酶一起在肌肉功能中起重要作用 将通过选择第二位点抑制子突变来鉴定。 导致共同本地化的发展组装过程 将对糖酵解酶进行表征。瞄准更多的 糖酵解酶将被研究，其中之一，醛缩酶，将是 广泛发展。这将需要隔离和表型 醛缩酶基因突变体的特性研究。C-的作用-- 肌醛缩醛酶肌特异性异构体的末端将通过 分析携带转基因的转基因果蝇的飞行情况。 最后，组装过程的细节将被描述为 利用分离的肌原纤维进行体外检测的进一步发展 作为一个平台，在其上分析 糖酵解复合体在Z-Disc/M-线上组装和组织。这个 我们的工作结果将对理解 碳水化合物分解代谢、能量产生与肌肉的关系 功能。这些影响很可能会延伸到 果蝇模型，应与理解缺陷相关 人体肌肉功能。

This research program has as its goal understanding the structural basis for glycolytic enzyme co-localization and targeting at Z-discs and M- lines of the sarcomere as a means of understanding how this essential aspect of glycolytic enzyme organization provides the energy for muscle contraction. In the flight muscle of Drosophila, the glycolytic enzymes, glycerol-3-phosphate dehydrogenase (GPDH), glyceraldehyde-3- phosphate dehydrogenase (GAPDH) and aldolase are localized at Z-discs/M- lines. Localization of GAPDH and aldolase depends on the simultaneous localization of GPDH. In the absence of glycolytic enzyme co- localization in flight muscles, flies can't fly. Further elucidation of the nature of protein-protein interactions that support colocalization will lead to a better understanding of the mechanisms by which ATP production from glycolysis supports muscle function. It remains to be determined whether glycolytic enzymes physically interact or whether other proteins play a role in co-localization. This will be determined using the yeast two hybrid system and co-immunoprecipitation techniques. The C-terminal tripeptide, Q-N, L found in the muscle specific isoform, GPDH-1 is required for GPDH targeting. Other domains of the glycolytic enzymes may play a role in targeting these enzymes to the Z-discs/M- lines and will be identified. Genes which encode proteins that interact with GPDH and aldolase and are functionally important in muscle function will be identified by selection of second site suppressor mutations. The developmental assembly process that results in co-localized glycolytic enzymes will be characterized. Targeting of additional glycolytic enzymes will be studied and one of these, aldolase, will be extensively developed. This will require isolation and phenotypic characterization of mutants in the aldolase gene. The role of the C- terminus in the aldolase muscle specific isoform will be evaluated by analyzing flight in transgenic flies which carry engineered transgenes. Finally, the details of the assembly process will be characterized by further development of an in vitro assay which uses isolated myofibrils as a platform on which to analyze the mechanism through which the glycolytic complex is assembled and organized at the Z-disc/M-line. The results of our work will have broad implications for understanding the connection between carbohydrate catabolism, energy production and muscle function. It is likely these implications will extend beyond the Drosophila model and should be relevant to understanding defects in human muscle function.