Structural dynamics of the 180 kDa HIV-1 initiation complex investigated using hydrogen/deuterium exchange mass spectrometry

摘要

We developed a new strategy for characterizing asymmetric heterodimers with shared sequence (e.g., reverse transcriptase (RT)) using HDX-MS. This was achieved by subunit-selective labeling of RT with ~(15)N to facilitate detection of peptides with identical sequences originating from either subunit based on their unique mass. Deuterium uptake following on-exchange was determined using a modified version of HDX Workbench (freely available upon request). Isotopic labeling of RT did not affect the intrinsic dynamics of the protein.