I am working with relatively smaller membrane protein of size 28kD.I recently purified it as well. Is it worth a shot to reassemble it in nanodisc for functional and structural studies? Do proteins have to be of certain size for them to be reconstituted in a nanodisc?
I assume, you mean lipid-protein nanodisc (LPN)? If yes, it is difficcult to say a priori, whether these nanodiscs are suitable to study your protein. However, successfull usage cases of LPN are reported (http://www.ncbi.nlm.nih.gov/pubmed/20356311). I would also start from reading some review papers on NMR applications of LPN (http://www.ncbi.nlm.nih.gov/pubmed/19747096). And the last question (about certain size): the answer is - yes. However, LPN differ in the type of apolipoprotein used for its rim stabilization. You have a choice of them to vary the size of the diameter of the flat part of the disc. Thus, the variation of the protein sizes suitablle for reconstitution into nanodisc can be enlarged.
Hi. your protein should fit into all the most commonly used Nanodiscs, eg using the MSPD1 (regular) or MSPD1E3 (enlarged). For functional assays the size of the disc probably doesn't have an influence, you might have more than one protein per disc in the bigger ones, though. If you are going for solution NMR spectroscopy, the disc should be as small as possible.
Hi Greta,
As Tobias said already, for functional studies size of your protein-ND complex doesn't matter. For structural studies, it really depends. I am not much aware of new developments in NMR but with 28kDa protein single particle cryo-em will still be very challenging. However, there are few cases where IMP were crystallised in NDs using LCP, even to 1.8 Å. If you are able to reconstitute the protein you that may be worth trying. If you have functional Fab against your protein that may be helpful in single particle analysis.
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