link http://precedings.nature.com/documents/4479/version/1/files/npre20104479-1.pdf doi:10.1038/npre.2010.4479.1 Osmotic stress and water isotope effects in kinesin-1 gliding motility assays Steven J. Koch1, Andy Maloney1 & Lawrence J. Herskowitz1 Correspondence: (Login to view email address) University of New Mexico, Dept. Physics and Astronomy and CHTM PPT (3.9 MB) PDF (1.9 MB) Download: PDF (1.9 MB) Embed: License: Document Type: Presentation Date: Received 21 May 2010 00:13 UTC; Posted 21 May 2010 Subjects: Molecular Cell Biology Tags: kinesin kinesin-1 gliding motility assay osmotic pressure Osmotic stress heavy water water isotope effects deuterium oxygen-18 heavy-oxygen water microtubules Abstract: The osmotic pressure and kinetic properties of water play important roles in biomolecular interactions. As pointed out by Parsegian, Rand, and Rau, these crucial roles are often overlooked1. In some fields, osmotic stress and isotope effects have been exploited for probing the role water plays in binding interactions of biomolecules. To our knowledge, there have been no studies of osmotic stress and water isotope effects for kinesin, and only a handful for myosin. We’re currently using the gliding motility assay to see whether we can extract new information about kinesin-1 / microtubule interactions by changing osmotic stress and water isotopes. We will describe our open-source, automated analysis platform for extracting microtubule gliding speeds from image series. We will also show our preliminary analyses of the changes seen in gliding assays when done in heavy water (either heavy-hydrogen or heavy-oxygen) or osmolytes (betaine). We will discuss whether osmotic stress and isotopes, particularly heavy-oxygen water, might be an important tool for probing effects of water on binding interactions between kinesin and microtubules. We will also discuss potential applications of deuterium water for stabilizing microtubules and kinesin for lab or device applications. [1] Parsegian, V. A., Rand, R. P., & Rau, D. C. (1995). Macromolecules and water: probing with osmotic stress. Methods in Enzymology, 259. This work was supported by the DTRA CB Basic Research Program under Grant No. HDTRA1-09-1-008 in collaboration with Dr. Susan Atlas lab (UNM). Presented at: Thermodynamics and Kinetics of Molecular Motors, 20 May 2010 Discussion Comments: 0 comments Share: (Login to share with a colleague) Additional information License: This document is licensed to the public under the Creative Commons Attribution 3.0 License How to cite this document: Koch, Steven, Maloney, Andy, and Herskowitz, Lawrence. Osmotic stress and water isotope effects in kinesin-1 gliding motility assays. Available from Nature Precedings (2010) Version info: Other versions of this document in Nature Precedings v3 Posted 09 March 2011 v2 Posted 01 June 2010 Other versions of this document elsewhere on the web http://www.slideshare.net/skoch3/osmotic-stress-and-water-isotope-effects-in-kinesin1-gliding-motility-assays : Includes the *.pptx file