Abstract
Bacteriophage T4 lysozyme (T4L) has been used as a paradigm for seminal biophysical studies on protein structure, dynamics, and stability. Approximately 700 mutants of this protein and their respective complexes have been characterized by X-ray crystallography; however, despite the high resolution diffraction limits attained in several studies, no hydrogen atoms were reported being visualized in the electron density maps. To address this, a 2.2 Å-resolution neutron data set was collected at 80 K from a crystal of perdeuterated T4L pseudo-wild type. We describe a near complete atomic structure of T4L, which includes the positions of 1737 hydrogen atoms determined by neutron crystallography. The cryogenic neutron model reveals explicit detail of the hydrogen bonding interactions in the protein, in addition to the protonation states of several important residues.
| Original language | English |
|---|---|
| Pages (from-to) | 2098-2104 |
| Number of pages | 7 |
| Journal | Protein Science |
| Volume | 26 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2017 |
Funding
This research was sponsored by the Laboratory Directed Research and Development Program of Oak Ridge National Laboratory (ORNL), managed by UT-Battelle, LLC, for the U.S. Department of Energy (DOE) under Contract No. DE-AC05-00OR22725. Research conducted at ORNL\u2019s High-Flux Isotope Reactor and Spallation Neutron Source was sponsored by the US Department of Energy\u2019s (DOE) Office of Basic Energy Sciences, Scientific User Facilities Division, and laboratory facilities supported by the DOE Office of Biological and Environmental Research, Project ERKP291. The IMAGINE Project was partially supported by the National Science Foundation (Grant 0922719).
Keywords
- T4-lysozyme
- X-ray
- hydrogen
- neutron
- structure