Plant & zo
The science of plants and more
Work in progress
This last week I have been busy with organising my results and preparing them for a presentation that I gave for some of our department. And although I have made some progress with my project since I started it, the presentation I gave highlighted the lack of useful progress during the last half year or so. I say useful progress on purpose. Of course you always make some progress, if only by figuring out why something did not work. But my last year has been basically that, finding out why something did not work in my protein purification procedure, amend this, just to have something else going wrong.
So what I try to do is purify proteins that bind to a phosphoinositol lipid. These phosphoinositol lipids come in different flavours which have both a specific shape and a charge. This enables proteins that bind those phosphoinositol lipids to discriminate between them. Although, you have proteins that bind depending both shape and charge, specific binders, however, others will bind based on charge only, unspecific binders. In my project we are primarily interested in those specific binders. In order to determine which proteins bind specifically and which unspecifically we like a step in the purification procedure that would allow us to discriminate between them. And it is step that primarily gives me trouble. Sometimes it works, but then at other times it doesn’t. As if having optimised the binding to the phosphoinositol-beads affects the ability to discriminate between specific and unspecific binding proteins.
The good thing that came out of my talk was that when another PI suggested to do the purification without the step to discriminate between specific and unspecific binders, in the end my PI gave in. I now will be faced with the larger task afterwards to weed out the unspecific from the specific binders after having identified the proteins from the purification. But for now I am excited to finally be able to do the big scale purification for the protein identification to get that list of phosphoinositide binders to work with.