Proteomics of Single Muscle Fibers!

There is a lot of basic physiology to learn from in this great new study study in "Histology." It's so cool to pick up study that you can already follow most of (in my case, the proteomics, most of the time) and then use that framework to fill in all sorts of things you'd never thought about at all.

You can access the preformatted text here (warning, direct download, if that is a problem. I'm having some trouble finding the appropriate Journal link on this tablet.)

How would you start doing some muscle proteomics? Before this popped into my Google feed I would have taken a sample and homogenized enough to digest.

You muddle all the information! There are different types of muscle fibers that are controlled by distinct genetic machinery. There are fast ones and slow ones and there are slow ones that fatigue slow and some fatigue fast, and while these sound like vague generalizations they're actually distinct tissues with their own proteomic and metabolomic characteristics!

Cool, right?

It is until you realize that an individual fiber may only contain 5 micrograms of protein! Digest and desalt that and you'd better not have a bubble in your NanoLC line. Wait! It's 2019! If we're smart about it, 5ug is a ton of material!

They combine some material to build their library and then utilize match between runs (MBR) to characterize the individual materials and obtain plenty of coverage. They find really interesting differences in the mitochondrial proteins of the individual cells of interest and it's enough for some solid conclusions about the muscle biology.