Intracellular protein degradation by the ubiquitin-proteasome system is ATP dependent, and the optimal ATP concentration to activate proteasome function in vitro is -100 μM. IntraceUular ATP levels are generally in t...Intracellular protein degradation by the ubiquitin-proteasome system is ATP dependent, and the optimal ATP concentration to activate proteasome function in vitro is -100 μM. IntraceUular ATP levels are generally in the low millimolar range, but ATP at a level within this range was shown to inhibit proteasome peptidase activities in vitro. Here, we report new evidence that supports a hypothesis that intracellular ATP at the physiological levels bidirectionally regulates 26S proteasome proteolytic function in the cell. First, we confirmed that ATP exerted bidirectional regulation on the 26S proteasome in vitro, with the optimal ATP concentration （between 50 and 100μM） stimulating proteasome chymotrypsin-like activities. Second, we found that manipulating intracellular ATP levels also led to bidirectional changes in the levels of proteasome-specific protein substrates in cultured cells. Finally, measures to increase intracellular ATP enhanced, while decreasing intraceHular ATP attenuated the ability of proteasome inhibition to induce cell death. These data strongly suggest that endogenous ATP within the physiological concentration range can exert a negative impact on proteasome activities, allowing the cell to rapidly upregulate proteasome activity on ATP reduction under stress conditions.展开更多
Object. The effects of ATP-introduced a rise in cytosolic free Ca2+ concentration and inhibition of nitric oxide were investigated. Method. Measurement of free Ca2+([Ca2+] i)of cultured rat tail arterial smooth muscle...Object. The effects of ATP-introduced a rise in cytosolic free Ca2+ concentration and inhibition of nitric oxide were investigated. Method. Measurement of free Ca2+([Ca2+] i)of cultured rat tail arterial smooth muscle cells using Fura-2/AM dual excitation wavelength spectrofluorometer. Results. There are two components of [Ca2+] i can be evoked by ATP. One part is Ca2+ entry from Ca2+ channel and formed a plateau. The another part is a peak that released from Ca2+ store. Both of them can be inhibited by NO. Conclusion. The ATP induced [Ca2+] i rise that release Ca2+ from both Insp 3 and ryanochine receptors and Ca2+ entry through calcium channels. The inhibition of NO on ATP induced [Ca2+] i rise that was mediated by cGMP.展开更多
F_1-ATPase, a part of ATP synthase, can synthesize and hydrolyze ATP moleculars in which the centralγ-subunit rotates inside the α_3β_3 cylinder.A stochastic four-state mechanochemical coupling model of F_1-ATPase ...F_1-ATPase, a part of ATP synthase, can synthesize and hydrolyze ATP moleculars in which the centralγ-subunit rotates inside the α_3β_3 cylinder.A stochastic four-state mechanochemical coupling model of F_1-ATPase isstudied with the aid of the master equation.In this model, the ATP hydrolysis and synthesis are dependent on ATP,ADP, and Pi concentrations.The effects of ATP concentration, ADP concentration, and the external torque on theoccupation probability of binding-state, the rotation rate and the diffusion coefficient of F_1-ATPase are investigated.Moreover, the results from this model are compared with experiments.The mechanochemical mechanism F_1-ATPase isqualitatively explained by the model.展开更多
基金Acknowledgments This work was supported by the National High Technol- ogy Research and Development Program of China （Project 2006AA02Z4B5）, the National Natural Science Foundation of China （Project 2010）, and a Key Project （9251018201002） of Guangdong Province Natural Science Foundation （to JL）. It was also supported in part by Grants HL072166, HL085629, and HL068936 of the NIH and an Established Investigator Award （0740025N） of the American Heart Association （to XW）.
文摘Intracellular protein degradation by the ubiquitin-proteasome system is ATP dependent, and the optimal ATP concentration to activate proteasome function in vitro is -100 μM. IntraceUular ATP levels are generally in the low millimolar range, but ATP at a level within this range was shown to inhibit proteasome peptidase activities in vitro. Here, we report new evidence that supports a hypothesis that intracellular ATP at the physiological levels bidirectionally regulates 26S proteasome proteolytic function in the cell. First, we confirmed that ATP exerted bidirectional regulation on the 26S proteasome in vitro, with the optimal ATP concentration （between 50 and 100μM） stimulating proteasome chymotrypsin-like activities. Second, we found that manipulating intracellular ATP levels also led to bidirectional changes in the levels of proteasome-specific protein substrates in cultured cells. Finally, measures to increase intracellular ATP enhanced, while decreasing intraceHular ATP attenuated the ability of proteasome inhibition to induce cell death. These data strongly suggest that endogenous ATP within the physiological concentration range can exert a negative impact on proteasome activities, allowing the cell to rapidly upregulate proteasome activity on ATP reduction under stress conditions.
文摘Object. The effects of ATP-introduced a rise in cytosolic free Ca2+ concentration and inhibition of nitric oxide were investigated. Method. Measurement of free Ca2+([Ca2+] i)of cultured rat tail arterial smooth muscle cells using Fura-2/AM dual excitation wavelength spectrofluorometer. Results. There are two components of [Ca2+] i can be evoked by ATP. One part is Ca2+ entry from Ca2+ channel and formed a plateau. The another part is a peak that released from Ca2+ store. Both of them can be inhibited by NO. Conclusion. The ATP induced [Ca2+] i rise that release Ca2+ from both Insp 3 and ryanochine receptors and Ca2+ entry through calcium channels. The inhibition of NO on ATP induced [Ca2+] i rise that was mediated by cGMP.
基金Supported by the National Natural Science Foundation of China under Grant No.10847118the National Natural Science Foundation of the City of Tianjin under Grant No.08JCYBJC00900 the Science Research Program of Education office of Hebei Province under Grant No.2008427
文摘F_1-ATPase, a part of ATP synthase, can synthesize and hydrolyze ATP moleculars in which the centralγ-subunit rotates inside the α_3β_3 cylinder.A stochastic four-state mechanochemical coupling model of F_1-ATPase isstudied with the aid of the master equation.In this model, the ATP hydrolysis and synthesis are dependent on ATP,ADP, and Pi concentrations.The effects of ATP concentration, ADP concentration, and the external torque on theoccupation probability of binding-state, the rotation rate and the diffusion coefficient of F_1-ATPase are investigated.Moreover, the results from this model are compared with experiments.The mechanochemical mechanism F_1-ATPase isqualitatively explained by the model.
