2LCB

Solution Structure of a Minor and Transiently Formed State of a T4 Lysozyme Mutant

Summary for 2LCB

• Entry DOI: 10.2210/pdb2lcb/pdb
• Related: 2LC9 3DMV 3DMX
• NMR Information: BMRB: 17604
• Descriptor: Lysozyme (1 entity in total)
• Functional Keywords: excited state, hydrolase
• Biological source: Enterobacteria phage T4
• Total number of polymer chains: 1
• Total formula weight: 18586.28

Authors

Bouvignies, G.,Vallurupalli, P.,Hansen, D.,Correia, B.,Lange, O.,Bah, A.,Vernon, R.M.,Dahlquist, F.W.,Baker, D.,Kay, L.E. (deposition date: 2011-04-26, release date: 2011-08-17, Last modification date: 2024-05-01)

Primary citation

Bouvignies, G.,Vallurupalli, P.,Hansen, D.F.,Correia, B.E.,Lange, O.,Bah, A.,Vernon, R.M.,Dahlquist, F.W.,Baker, D.,Kay, L.E.
Solution structure of a minor and transiently formed state of a T4 lysozyme mutant.
Nature, 477:111-114, 2011

PubMed Abstract: Proteins are inherently plastic molecules, whose function often critically depends on excursions between different molecular conformations (conformers). However, a rigorous understanding of the relation between a protein's structure, dynamics and function remains elusive. This is because many of the conformers on its energy landscape are only transiently formed and marginally populated (less than a few per cent of the total number of molecules), so that they cannot be individually characterized by most biophysical tools. Here we study a lysozyme mutant from phage T4 that binds hydrophobic molecules and populates an excited state transiently (about 1 ms) to about 3% at 25 °C (ref. 5). We show that such binding occurs only via the ground state, and present the atomic-level model of the 'invisible', excited state obtained using a combined strategy of relaxation-dispersion NMR (ref. 6) and CS-Rosetta model building that rationalizes this observation. The model was tested using structure-based design calculations identifying point mutants predicted to stabilize the excited state relative to the ground state. In this way a pair of mutations were introduced, inverting the relative populations of the ground and excited states and altering function. Our results suggest a mechanism for the evolution of a protein's function by changing the delicate balance between the states on its energy landscape. More generally, they show that our approach can generate and validate models of excited protein states.

PubMed: 21857680
DOI: 10.1038/nature10349

Experimental method

SOLUTION NMR