Protein structures are dynamic, undergoing motions that can play a vital role in function. However, the link between primary sequence and conformational dynamics remains poorly understood. Here, we studied how conformational dynamics can arise in a globular protein by evaluating the impact of individual core-residue substitutions in DANCER-3, a streptococcal protein G domain β1 variant that we previously designed to undergo a specific mode of conformational exchange that has never been observed in the wild-type protein. Using a combination of solution NMR experiments and molecular dynamics simulations, we demonstrate that only two mutations are necessary to create this conformational exchange, and that these mutations work synergistically, with one destabilizing the native structure and the other allowing two new conformational states to be accessed on the energy landscape. Overall, our results show how dynamics can appear in a stable globular fold, a critical step in the molecular evolution of dynamics-linked functions.
Submitter: Marc Mayer
Submission Date: May 9, 2020, 11:10 a.m.
This is an updated version of study HciYX5Lf3.
|Number of data points||54|
|Proteins||Immunoglobulin G-binding protein G|
|Assays/Quantities/Protocols||Experimental Assay: Protein G(B1) unfolding free energy ; Experimental Assay: Protein G(B1) oligomerization ; Experimental Assay: Protein G (B1) chemical stability ; Experimental Assay: Protein G (B1) melting temperature ; Derived Quantity: SD of Protein G (B1) chemical stability ; Derived Quantity: SD of Protein G(B1) unfolding free energy|
|Libraries||Stability and Dynamicity of GB1 Variants|
|Structure ID||Release Date||Resolution||Structure Title|
|1EM7||2000-03-16T00:00:00+0000||2.0||HELIX VARIANT OF THE B1 DOMAIN FROM STREPTOCOCCAL PROTEIN G|
|1GB1||1991-05-15T00:00:00+0000||0||A NOVEL, HIGHLY STABLE FOLD OF THE IMMUNOGLOBULIN BINDING DOMAIN OF STREPTOCOCCAL PROTEIN G|
|1IGC||1994-08-05T00:00:00+0000||2.6||IGG1 FAB FRAGMENT (MOPC21) COMPLEX WITH DOMAIN III OF PROTEIN G FROM STREPTOCOCCUS|
|1IGD||1994-08-05T00:00:00+0000||1.1||THE THIRD IGG-BINDING DOMAIN FROM STREPTOCOCCAL PROTEIN G: AN ANALYSIS BY X-RAY CRYSTALLOGRAPHY OF THE STRUCTURE ALONE AND IN A COMPLEX WITH FAB|
|1LE3||2002-04-09T00:00:00+0000||0||NMR Structure of Tryptophan Zipper 4: A Stable Beta-Hairpin Peptide Based on the C-terminal Hairpin of the B1 Domain of Protein G|
|1MPE||2002-09-12T00:00:00+0000||0||Ensemble of 20 structures of the tetrameric mutant of the B1 domain of streptococcal protein G|
|1MVK||2002-09-25T00:00:00+0000||2.5||X-ray structure of the tetrameric mutant of the B1 domain of streptococcal protein G|
|1PGA||1993-11-23T00:00:00+0000||2.07||TWO CRYSTAL STRUCTURES OF THE B1 IMMUNOGLOBULIN-BINDING DOMAIN OF STREPTOCOCCAL PROTEIN G AND COMPARISON WITH NMR|
|1PGB||1993-11-23T00:00:00+0000||1.92||TWO CRYSTAL STRUCTURES OF THE B1 IMMUNOGLOBULIN-BINDING DOMAIN OF STREPTOCCOCAL PROTEIN G AND COMPARISON WITH NMR|
|1PGX||1992-04-03T00:00:00+0000||1.66||THE 1.66 ANGSTROMS X-RAY STRUCTURE OF THE B2 IMMUNOGLOBULIN-BINDING DOMAIN OF STREPTOCOCCAL PROTEIN G AND COMPARISON TO THE NMR STRUCTURE OF THE B1 DOMAIN|