The role of tropomyosin phosphorylation in muscle.

原肌球蛋白磷酸化在肌肉中的作用。

• 批准号: 6994009
• 负责人: Brandon J Biesiadecki
• 金额: $ 4.83万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-16 至 2007-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6994009
• 关键词: SDS polyacrylamide gel electrophoresis; actins; calcium flux; cardiac myocytes; densitometry; gelsolin; heart contraction; laboratory mouse; matrix assisted laser desorption ionization; microfilaments; muscle metabolism; myocardial ischemia /hypoxia; myocardium; myosins; papillary muscles; phosphoproteins; phosphorylation; postdoctoral investigator; posttranslational modifications; protein binding; protein protein interaction; tropomyosin; two dimensional gel electrophoresis

DESCRIPTION (provided by applicant): The experiments proposed in this study investigate the hypothesis that tropomyosin phosphorylation (Tm-P) in the heart is altered by cardiac stress and this charge modification of Tm alters it's interactions within the thin filament to affect contraction. To date the modification of Tm resulting from cardiac stress and its effect on contraction are unknown. We propose three aims: I) To evaluate Tm-P in the stressed heart. Tm-P will be characterized in isolated perfused hearts treated with ischemia/reperfusion by phosphoprotein specific staining of 2-D gels and mass spectrometry. II) To determine the effect of Tm-P on binding to actin, troponin and to actin in the presence of troponin. Tm binding will be carried out by co-sedimentation and quantified on 2-D gels with densitometry. Ill) To determine the effect of Tm-P on the development of force in cardiac muscle. Calcium activated tension will be measured in skinned papillary bundles treated with gelsolin to selectively remove the thin filament and reconstituted with actin, troponin and non-phosphorylated or phosphorylated Tm. The knowledge gained from these experiments will provide novel insight into the regulatory mechanism of Tm-P and its effect on cardiac muscle contraction in health and disease.

描述（由申请人提供）：本研究提出的实验调查了以下假设：心脏中心中的心脏磷酸化（TM-P）会因心脏应激而改变，而TM的这种电荷改变会改变其在薄丝中的相互作用以影响收缩。迄今为止，由于心脏应力及其对收缩的影响引起的TM修饰尚不清楚。我们提出三个目标：i）评估压力心脏中的TM-P。 TM-P将以二-D凝胶和质谱法的磷酸蛋白特异性染色治疗的孤立灌注心脏进行表征。 ii）确定TM-P对肌动蛋白，肌钙蛋白和肌动蛋白的结合的影响。 TM结合将通过共同补习进行，并用光密度测定法对2-D凝胶进行定量。生病）确定TM-P对心肌力中力的发展的影响。钙活化的张力将在用凝胶素处理的皮肤乳头状束中测量，以选择性去除细丝，并用肌动蛋白，肌钙蛋白和非磷酸化或磷酸化的TM重构。从这些实验中获得的知识将为TM-P的调节机制及其对健康和疾病中心肌收缩的影响提供新的见解。