Abstract
A new method for the measurement of phosphate release in contracting and relaxed permeabilized muscle fibers is described. The assay is based on a genetically engineered phosphate-binding protein labeled with a coumarin fluorescent probe, which binds inorganic phosphate tightly and shows a fourfold increase in fluorescence upon binding. Measurements of Pi release on the millisecond time scale with sensitivity in the 10 microM range are obtained that provide new information about the relationship between ATP hydrolysis and force production.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brune M., Hunter J. L., Corrie J. E., Webb M. R. Direct, real-time measurement of rapid inorganic phosphate release using a novel fluorescent probe and its application to actomyosin subfragment 1 ATPase. Biochemistry. 1994 Jul 12;33(27):8262–8271. doi: 10.1021/bi00193a013. [DOI] [PubMed] [Google Scholar]
- Potma E. J., Stienen G. J., Barends J. P., Elzinga G. Myofibrillar ATPase activity and mechanical performance of skinned fibres from rabbit psoas muscle. J Physiol. 1994 Jan 15;474(2):303–317. doi: 10.1113/jphysiol.1994.sp020023. [DOI] [PMC free article] [PubMed] [Google Scholar]