Spectroscopic glimpses of the transition state of ATP hydrolysis trapped in a bacterial DnaB helicase.

ABSTRACT: The ATP hydrolysis transition state of motor proteins is a weakly populated protein state that can be stabilized and investigated by replacing ATP with chemical mimics. We present atomic-level structural and dynamic insights on a state created by ADP aluminum fluoride binding to the bacterial DnaB helicase from Helicobacter pylori. We determined the positioning of the metal ion cofactor within the active site using electron paramagnetic resonance, and identified the protein protons coordinating to the phosphate groups of ADP and DNA using proton-detected ³¹P,¹H solid-state nuclear magnetic resonance spectroscopy at fast magic-angle spinning > 100 kHz, as well as temperature-dependent proton chemical-shift values to prove their engagements in hydrogen bonds. ¹⁹

SUBMITTER: Malar AA 

PROVIDER: S-EPMC8421360 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature