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Significance Paramagnetic NMR spectroscopy is exquisitely sensitive for sparsely populated states in protein–protein interactions, and thus, it can provide important information on how protein–protein complexes form and evolve toward their productive state. However, the description of ensembles of protein–protein orientations is nontrivial, and great care must be taken when deriving biologically relevant results. We have applied an algorithm that restricts the conformational space sampled by the two partners to the maximum allowed for by the data. These ensembles can then be reduced assuming the principle of scarcity. We found that some states are linked to the main state through electrostatic pathways. Such paths help to identify those minor states that are able to evolve into the productive complex.

Identification of productive and futile encounters in an electron transfer protein complex