INTERACTION OF PYRUVATE KINASE ISOZYMES WITH ACTIN-CONTAINING FILAMENTS

丙酮酸激酶是控制该速率的关键酶之一 糖酵解和糖异生。 因此，它可能会关闭 代谢调节。 调查人员建议调查 丙酮酸激酶同工酶的酶活性是否能 通过与肌动蛋白丝的相互作用来控制。 磷酸化的作用也将影响它们的相互作用。 阐明。 这些工作人员将研究肌肉之间的相互作用 具有肌肉F-肌动蛋白和重组瘦蛋白同工酶（M1型） 丝，酵母和肝同工酶（L型）与酵母和 肝F-肌动蛋白。 相关代谢物对 还将研究相互作用。 的酶学性质 将肌动蛋白结合的酶与游离的酶进行比较， 酵素 在L型丙酮酸激酶的情况下， 将使用磷酸化和去磷酸化的酶。 的 丙酮酸激酶与肌动蛋白丝相互作用 将通过分离游离和肌动蛋白结合的酶来进行 使用超离心，并测定游离 酶在上清液中。 自由和肌动蛋白的动力学- 将使用pH stat研究结合的酶。 长期 我们研究的目的是研究改变 环境和细胞骨架成分对细胞生长的调节作用 细胞质酶的催化活性。

Pyruvate kinase is one of the key enzymes in controlling the rate of glycolysis and gluconeogenesis. As such, it is subject to close metabolic regulation. The investigators propose to investigate whether the enzymatic activity of pyruvate kinase isozyme could be controlled by their interaction with actin-containing filaments. The role of phosphorylation of their interaction will also be elucidated. These workers will study the interaction of muscle isozyme (type M1) with muscle F-actin and reconstituted thin filaments, the yeast and liver isozyme (type L) with yeast and liver F-actin, respectively. The effect of relevant metabolites on the interactions will also be studied. The enzymatic properties of the actin-bound enzyme will be compared with those of the free enzyme. In the case of type L pyruvate kinase, both the phosphorylated and dephosphorylated enzymes will be used. The interaction of pyruvate kinase with actin-containing filaments will be performed by separating the free and actin-bound enzyme using ultra-centrifugation, and assaying the activity of the free enzyme in the supernatant. The kinetics of the free and actin- bound enzyme will be studied using pH stat. The long term objective of our research is to study the effect of changing the environment and the cytoskeletal components on the regulation of the catalytic activity of cytoplasmic enzymes.