REGULATION OF ENERGY TRANSDUCTION IN SMOOTH MUSCLE
平滑肌能量传导的调节
基本信息
- 批准号:2392719
- 负责人:
- 金额:$ 35.71万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1994
- 资助国家:美国
- 起止时间:1994-04-06 至 1999-03-31
- 项目状态:已结题
- 来源:
- 关键词:adenosine diphosphate adenosine triphosphate chemical kinetics computer simulation enzyme activity flash photolysis high performance liquid chromatography intermolecular interaction laboratory rabbit muscle contraction muscle metabolism myosin light chain kinase myosins phosphoprotein phosphatase phosphoproteins phosphorylation portal vein vascular smooth muscle
项目摘要
The overall goal of this proposal is to elucidate the mechanism(s)
responsible for regulation of contractile protein interaction and the
resulting mechanical output in normally functioning smooth muscle. The
mechanism of regulation of smooth muscle is not fully understood, nor is
it known how this process is modified in pathological conditions such as
hypertension and asthma. Such information on normal function will provide
the framework for future studies on the etiology of altered smooth muscle
function as well as therapeutic strategies. The specific questions deal
with how phosphorylation of the myosin light chain regulates the number of
myosin molecules which are activated as well as the cycling rate and force
output from an activated myosin crossbridge. The first specific aim is to
determine whether there are different pools of activated crossbridges with
different cycling rates, and, if so, how myosin light chain
phosphorylation affects the distribution and the cycling rate of each
pool. How myosin light chain phosphorylation mediates an increase in the
cycling rate of unphosphorylated myosin will be determined. Isometric
conditions and mechanical perturbations such as shortening and stretching
will be studied. This will give insight into how the kinetics of
crossbridge cycling are controlled by the mechanical state of the muscle,
as well as the effects of smooth muscle's unique regulatory system on this
process. Other experiments will specifically test whether the degree of
Cooperative activation of unphosphorylated myosin can be modulated by
receptor activation, and whether the known activation mechanisms can
account for the regulation of number of crossbridges cycling and their
cycling rate during calcium-mediated contractions. The main experimental
approach 'will be a single turnover protocol to determine the rate at
which ADP bound to myosin is replaced with 3H-ADP from splitting of 3H-ATP
formed by photolysis of "caged" 3H-ATP. The second aim is to quantitate
the interaction of myosin light chain kinase and myosin light chain
phosphatase with the crossbridge cycle in smooth muscle. The action of
these enzymes on intermediates in the crossbridge cycle may alter the
kinetics of the completion of the cycle. A method will be developed to
permit measurement of the rate of phosphate turnover in the myosin light
chain. This will involve measurement of the rate at which 33P-phosphate
from gamma33P-ATP (derived from photolysis of caged gamma33P-ATP) appears
in the light chain. The experiments will give a quantitative assessment of
the rates of kinase and phosphatase activities compared to the kinetics of
the crossbridge cycle. The final specific aim will determine how the
ATPase of smooth muscle is controlled by mechanical conditions.
Specifically, ATPase measurements will be made during brief shortenings
and stretches and will be compared to isometric conditions. The
experiments will allow determination of the relationship between ATP
utilization by myosin and work output from the muscle, and will provide
evidence as to whether the variation in velocity of shortening under
different conditions results from an internal loading of fast cycling
crossbridges by those with slower cycling rates.
本提案的总体目标是阐明机制。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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THOMAS M BUTLER其他文献
THOMAS M BUTLER的其他文献
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{{ truncateString('THOMAS M BUTLER', 18)}}的其他基金
REGULATION OF ENERGY TRANSDUCTION IN SMOOTH MUSCLE
平滑肌能量传导的调节
- 批准号:
2226833 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
REGULATION OF ENERGY TRANSDUCTION IN SMOOTH MUSCLE
平滑肌能量传导的调节
- 批准号:
2226831 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
CATCH: Mechanochemistry and Regulation in Smooth muscle
CATCH:平滑肌的机械化学和调节
- 批准号:
6820934 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
REGULATION OF ENERGY TRANSDUCTION IN SMOOTH MUSCLE
平滑肌能量传导的调节
- 批准号:
2685409 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
CATCH: Mechanochemistry and Regulation in Smooth muscle
CATCH:平滑肌的机械化学和调节
- 批准号:
7268826 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
CATCH: Mechanochemistry and Regulation in Smooth muscle
CATCH:平滑肌的机械化学和调节
- 批准号:
7477799 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
CATCH: Mechanochemistry and Regulation in Smooth muscle
CATCH:平滑肌的机械化学和调节
- 批准号:
6938514 - 财政年份:1994
- 资助金额:
$ 35.71万 - 项目类别:
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