Communications Biology | May 12, 2021
Niloofar Abolhasani Khaje, Alexander Eletsky, Sarah E Beihn, Charles K Mobley, Monique J Rogals, Yoonkyoo Kim, Sushil K Mishra, Robert J Doerksen, Steffen Lindert, James H Prestegard, and Joshua S Sharp
Commun Biol. 2022 May 12;5(1):452. doi: 10.1038/s42003-022-03411-y.
Abstract
High resolution hydroxyl radical protein footprinting (HR-HRPF) is a mass spectrometry-based method that measures the solvent exposure of multiple amino acids in a single experiment, offering constraints for experimentally informed computational modeling. HR-HRPF-based modeling has previously been used to accurately model the structure of proteins of known structure, but the technique has never been used to determine the structure of a protein of unknown structure. Here, we present the use of HR-HRPF-based modeling to determine the structure of the Ig-like domain of NRG1, a protein with no close homolog of known structure. Independent determination of the protein structure by both HR-HRPF-based modeling and heteronuclear NMR was carried out, with results compared only after both processes were complete. The HR-HRPF-based model was highly similar to the lowest energy NMR model, with a backbone RMSD of 1.6 Å. To our knowledge, this is the first use of HR-HRPF-based modeling to determine a previously uncharacterized protein structure.
