Dear collegues,
I have a technical question about cell membrane study. I'm working on a transmembrane protein that belong to tetraspanins superfamily and I know that tetraspanins directly interact with cholesterol. I'm searching for a method (or collaborators) to study if the absence of tetraspanins could affect in some way the plasma membrane composition and structure (and also its physical characteristics). Do you have any suggestion?
Thanks in advance for helping me.
Cheers
Luca
Infrared (IR) spectroscopy is a useful technique in the study of membrane protein conformation and dynamics
Article Infrared spectroscopy of proteins
Hi Sebastian Schmitt
and Antonina Kokisheva , I'm not in the field but I'm going to read something to become more confident with this argument. Nice suggestions to study conformation and dynalics of membranes but what about cholesterol content? I was thinking about mass spec, but maybe the readout will be on the total content of cholesterol and not specifically on the membrane bilayer.Thanks for your useful answer.
Cheers
Luca
It of course depends a bit on the cells you are using, but it is possible to prepare plasma membrane ghosts from some cell types other than RBCs. I think I have seen paprrs on lymphocyte ghosts, at least, but these papers are from 1970s when this was a more popular approach. So perhaps you could isolate the plasma membrane for lipidomic analysis or just cholesterol analysis by mass spectrometry, although the problem may be the relatively large quantity of lipids required, but, again, depends on your system.
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