User:Andy Maloney/Kinesin & Microtubule Page/Osmotic stress/Osmolytes

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Motivation

The specific motivation behind looking at how kinesin and microtubules interact when the solvent they are in is stressed with osmolytes is to investigate the kinetics of the kinesin and microtubule system. For instance, I can investigate how water hydrates the proteins and how that hydration affects the speed of microtubules in a gliding motility assay.

Experiments

As with all gliding motility experiments, I will passivate the glass surface with casein. More specifically with whole casein from Sigma (C7078).

The experiments I plan to work on to investigate osmolytes and the kinesin+microtubule system include the following. There are some very important things to take into consideration when investigating the following experiments. As I obtain more knowledge on the subject of osmolytes, this list will increase. As for now, what Dr. Koch and I have discussed include:

  • What is the osmotic pressure of the solvent without the kinesin+microtubule system for each of the osmolytes?
    • For now, without a working osmometer I will have to rely on literature for this answer.
    • Steve Koch 02:29, 25 March 2010 (EDT): Evan did bring the vapor pressure osmometer. I think a junior student should get it working after this semester. Who? I don't know, since Brian is probably going to an REU.
  • How does the viscosity of the solvent change when adding the osmolytes?
    • Again, I will have to rely on literature for this answer. Luckily I have chosen very well known osmolytes to start this investigation with. So, I'm hoping that the information exists in the literature. Thanks go to BPS for allowing me to talk with other scientists on this subject and to find out what osmolytes can be used in cell systems.
      • Viscometer
      • I can also use a glass viscometer but, this may take some time to learn how to use.
  • Does the pH change? If so, I need to monitor it or at least get it back to the pH 6.89 I use.

Betain

I plan on investigating the speed of microtubules in a gliding motility assay with the following concentrations of betain.

  • 0 - 1000 uM

D-Proline

  • 0 - 1000 uM

Sarcosine

  • 0 - 1000 uM