User:Andy Maloney/Kinesin & Microtubule Page/Kinesin papers/Effect of temperature on kinesin driven microtubule gliding and kinesin ATPase activity
These are my notes on the below paper. Please do not read my notes until you have read the paper.
- This paper talks about the changes in gliding motility speeds due to temperature effects.
- They used porcine brain kinesin that they purified themselves.
- They purified tubulin from porcine brains as well. They polymerized the microtubules in 10 µM Taxol.
- Their buffer consisted of:
- 50 mM imidazole
- 0.5 mM MgCl2
- 0.5 mM EGTA
- 0.1 mM DTT
- pH 6.8
- They added 100 mM NaCl, 0.5 mM disodium ATP, and 10 µM Taxol for motility.
- Their final concentration of tubulin and kinesin were:
- Kinesin 70 µg/mL
- Tubulin 40 µg/mL
- They incubated microtubules and kinesin for 10 minutes before adding them to a slide.
- They used 5 mg/mL BSA as their passivator.
- They used the Argus 20 image processing software.
- The take home from this paper is that speed from gliding motility assays are dependent on the temperature.
- Steve Koch 23:34, 3 March 2010 (EST): One interesting thing I noticed upon re-reading. At Sandia we discovered that not only did thermomyces kinesin aggregate at 42C, but so did drosophila kinesin. Given the unique way in which they prepared their assays, I think they incubated the kinesin at high temperature in solution before putting on surface (Figure 3) ... so aggregation of kinesin likely accounts for the inactivity. As for figure 1, I don't know if the high temp behavior (plateau) is explained by the same thing.