I studied kinesin aggregation a bit when a postdoc at Sandia. See: http://openwetware.org/wiki/User:Steven_J._Koch/Kinesin_Aggregation
4:49 PM Thursday, February 12, 2004
Tried DIC microscopy on the aggregated kinesin today. Here are the raw notes:
Comparing 45C heated kin797h ("elution #2" from July 2003) with non-heated. (See DLS data for the heating.)
Both samples have "gook." But the gook is much more extensive in the heated sample. This could corroborate that the aggregation is the mechanism for inactivation, even when not heated. For the second set of images, (about 4:30 PM), I have waited about 5 minutes after loading the samples, in order to have comparable settling of heavy material.
-> "45C Aggregates Image Series" is from the first aggregates sample. To give an idea of what the sample looks like...focusing up and down as well.
-> 01 photo is the first 45C sample.
-> 02-04 photos are the second 45C sample, on the dual (comparison) slide
-> 05-07 photos are the 25C (control) sample. You can see the similar aggregates, but at a much lower density
Here is a sample image:
I am running out of time before leaving for the meeting, but here are the general thoughts:
The aggregates that I can see don’t strike me as “amyloid.” However, they aren’t quite “globular” either, and you get the impression that they are like 1-D strings (polymers).
There are more aggregates in the heated sample
Indicating that this “junk” is the kin797H aggregates.
The aggregates are similar in both the heated and unheated samples
Indicating that some aggregation is occurring even at 4C.
And that this is perhaps the mechanism for inactivation during dormancy
Perhaps I don’t really need EM to visualize these things?