Structure Determination Protocol for Transmembrane Domain Oligomers

. Author manuscript; available in PMC: 2020 May 20.

Published in final edited form as: Nat Protoc. 2019 Jul 3;14(8):2483–2520. doi: 10.1038/s41596-019-0188-9

Step	Problem	Possible reason	Possible solution
11	No inter-NOEs are observed	The ²H,¹⁵N- and ¹³C-label proteins have not been mixed adequately	Mix the two preparations before performing CNBr cleavage
	Positive inter-NOEs are present in the sub-spectrum A, but negative inter-NOEs are weak or not present in the sub-spectrum B	NMR relaxation is more pronounced in the pathway B of the experiment	Perform the analysis as described. Inter-NOEs in the inter-NOEs selective spectrum (A-B) may appear weaker, but intra-NOEs will still be completely canceled