Author:
Malär Alexander A.,Wili Nino,Völker Laura A.,Kozlova Maria I.,Cadalbert Riccardo,Däpp Alexander,Weber Marco E.,Zehnder Johannes,Jeschke Gunnar,Eckert Hellmut,Böckmann Anja,Klose Daniel,Mulkidjanian Armen Y.,Meier Beat H.,Wiegand Thomas
Abstract
AbstractThe 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 fromHelicobacter 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-detected31P,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.19F and27Al MAS NMR spectra reveal a highly mobile, fast-rotating aluminum fluoride unit pointing to the capture of a late ATP hydrolysis translation state in which the phosphoryl unit is already detached from the arginine and lysine fingers.
Publisher
Cold Spring Harbor Laboratory