Site-Selective Dynamics of Ligand-Free and Ligand-Bound Azidolysozyme

TL;DR

This study demonstrates that azido-modified alanine residues serve as environment-sensitive probes capable of detecting ligand binding and revealing protein dynamics through changes in vibrational frequencies and correlation functions.

Contribution

The paper introduces the use of azido-modified alanine residues as versatile probes for site-specific investigation of protein structure and dynamics, especially in relation to ligand binding.

Findings

Distinct vibrational frequency shifts upon ligand binding

Changes in long-time decay and static offset of frequency fluctuations

Mapping of dynamics onto residue-residue coupling via cross-correlation maps

Abstract

Azido-modified alanine residues (AlaN$_3$) are environment-sensitive, minimally invasive infrared probes for the site-specific investigation of protein structure and dynamics. Here, the capability of the label is investigated to query whether or not a ligand is bound to the active site of Lysozyme and how the spectroscopy and dynamics change upon ligand binding. The results demonstrate specific differences for center frequencies of the asymmetric azide stretch vibration, the long time decay and the static offset of the frequency fluctuation correlation function - all of which are experimental observables - between the ligand-free and the ligand-bound, N$_3$-labelled protein. Changes in dynamics can also be mapped onto changes in the local and through-space coupling between residues by virtue of dynamical cross-correlation maps. This makes the azide label a versatile and structurally sensitive probe to report on the dynamics of proteins in a variety of environments and for a range of different applications.