Core--Chemical

化学核心的长期目标和具体目标是提供独特的荧光和光敏“笼式”核苷酸资源，用于探索平滑肌生理学和细胞生物学的机制。与该方案项目相关的研究的具体健康益处列于导言(意义“和个别项目中)。Core将主要通过(1)合成香豆素标记的核苷酸，从而能够识别交叉桥状态并测量其形成和衰退的速度；(2)合成新型的笼状核苷酸和包括笼状磷酸化肌球蛋白调节轻链在内的平滑肌调节器，从而实现其目标。实现这些目标的方法将是通过在主要调查人员的实验室中制定的化学方案。这一合成化学计划将得到对探针(通过监测它们与分离的肌肉蛋白相互作用时的活性和功能)和笼状化合物(通过监测它们的光化学性质和(理想的)生物惰性)的严格表征的支持。进一步的目标将通过开发PI和ADP的实时探针来实现，这些探针将被应用(项目1)来记录不同生理状态下的平滑肌ATPase活性和伪ATPase(肌球蛋白轻链激酶/平滑肌磷酸酶)活性。最后，只要可行，这些试剂将在生理环境中通过探测它们在去皮骨骼肌纤维中的行为来进行测试。NIMR物理生物化学部的化学家、酶学家和生理学家的专业知识和合作使其成为一个独特的化学核心设施。

The Chemical Core has the long term objective and specific aim of providing a unique resource of fluorescent and photosensitive "caged" nucleotides for exploring mechanisms of smooth muscle physiology and cell biology. The specific health benefits of the research relevant to this program project are set out in the Introduction (Significance" and in the individual projects. The Core will achieve its goals principally through (1) synthesis of coumarin-labeled nucleotides that will enable crossbridge states to be identified and their rates of formation and decay measured; (2) the synthesis of novel caged nucleotides and regulators of smooth muscle including caged phosphorylated myosin regulatory light chains. The methods for achieving these goals will be through chemical protocols established in the laboratories of the principal investigators. This synthetic chemistry program will be supported by rigorous characterization of the probes (by monitoring their activity and function when interacting with isolated muscle proteins) and the caged compounds (by monitoring their photochemical properties and (desirable) biological inertness). Further goals will be achieved by development of real-time probes of Pi and ADP that will be applied (Project 1) to record smooth muscle ATPase activity in various physiological states and pseudo- ATPase (myosin light chain kinase/smooth muscle phosphatase) activity. Finally, the reagents will, insofar as is practical, be tested in a physiological environment by probing their behavior in skinned skeletal muscle fibers. The expertise and collaboration of chemists, enzymologists and physiologists in the Division of Physical Biochemistry at NIMR make this a unique chemical core facility.