期刊
JOURNAL OF GENERAL PHYSIOLOGY
卷 151, 期 6, 页码 860-869出版社
ROCKEFELLER UNIV PRESS
DOI: 10.1085/jgp.201812243
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资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan [22227005, 23107003, 15H04677, 18K06878, 26560225, 15K12524, 25707035, 16K18993]
- Japan Society for the Promotion of Science [JP15J10205]
- PRESTO, Japan Science and Technology Agency [JPMJPR17P3, JPMJPR15F5]
- Japan Heart Foundation
- Grants-in-Aid for Scientific Research [15H04677, 18K06878, 26560225, 25707035, 16K18993, 15K12524, 22227005] Funding Source: KAKEN
During the excitation-contraction coupling of the heart, sarcomeres are activated via thin filament structural changes (i.e., from the off state to the on state) in response to a release of Ca2+ from the sarcoplasmic reticulum. This process involves chemical reactions that are highly dependent on ambient temperature; for example, catalytic activity of the actomyosin ATPase rises with increasing temperature. Here, we investigate the effects of rapid heating by focused infrared (IR) laser irradiation on the sliding of thin filaments reconstituted with human a-tropomyosin and bovine ventricular troponin in an in vitro motility assay. We perform high-precision analyses measuring temperature by the fluorescence intensity of rhodamine-phalloidin-labeled F-actin coupled with a fluorescent thermosensor sheet containing the temperature-sensitive dye Europium (III) thenoyltrifluoroacetonate trihydrate. This approach enables a shift in temperature from 25 degrees C to similar to 46 degrees C within 0.2 s. We find that in the absence of Ca2+ and presence of ATP, IR laser irradiation elicits sliding movements of reconstituted thin filaments with a sliding velocity that increases as a function of temperature. The heating-induced acceleration of thin filament sliding likewise occurs in the presence of Ca2+ and ATP; however, the temperature dependence is more than twofold less pronounced. These findings could indicate that in the mammalian heart, the on-off equilibrium of the cardiac thin filament state is partially shifted toward the on state in diastole at physiological body temperature, enabling rapid and efficient myocardial dynamics in systole.
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