Abstract
Deletion of the protein-protein interaction SH3 domain of the membrane remodeling amphiphysin 2 (BIN1) protein was found to lead to centronuclear myopathy in patients, yet only few interaction partners of BIN1 SH3 have been identified so far, precluding a better understanding of the pathomechanism. Here we used the holdup assay to proteome-wide measure steady-state affinity constants of BIN1 SH3 domain for thousands of full-length cellular proteins, as well as for hundreds of putative SH3-binding sites found within the identified BIN1 partners. Besides confirming known partners, such as dynamin 2 (DNM2), we also identified and affinity-characterized numerous others, like SMCHD1, which were previously implicated in different neuromuscular disorders. We also assessed the impact of a set of rare natural BIN1 SH3 domain variants on affinity interactomes and identified potentially harmful ones that exhibited perturbed affinity profiles, whose impacts were confirmed in a cellular assay for BIN1-mediated membrane remodeling, tentatively connecting them to neuromuscular disorders. In the study, we develop a new affinity-interactomic strategy, which can be generally applied to study the consequences of disease-associated genomic variants of any kind.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
This revised preprint contains minor changes in the title, main text, as well as a change in the order of the author list compared to the original submission.