The evolution of proteins with novel function is thought to start from precursor proteins that are conformationally heterogeneous. The corresponding genes may be duplicated and then mutated to select and optimize a specific conformation. However, testing this idea has been difficult because of the challenge of quantifying protein flexibility and conformational heterogeneity as a function of evolution. Here, we report the characterization of protein heterogeneity and dynamics as a function of evolution for the antifluorescein antibody 4-4-20. Using nonlinear laser spectroscopy, surface plasmon resonance, and molecular dynamics simulations, we demonstrate that evolution localized the Ab-combining site from a heterogeneous ensemble of conformations to a single conformation by introducing mutations that act cooperatively and over significant distances to rigidify the protein. This study demonstrates how protein dynamics may be tailored by evolution and has important implications for our understanding of how novel protein functions are evolved.
ID: pPcLSu4G
Submitter: Stephanie Contreras
Submission Date: June 26, 2020, 10:36 a.m.
Version: 1
Number of data points | 132 |
Proteins | VL(4-4-20) ; VH(4-4-20) ; VL(gl) ; Ig gamma-2A chain C region, membrane-bound form,4-4-20 (IG*G2A=KAPPA=) FAB FRAGMENT |
Unique complexes | 22 |
Assays/Quantities/Protocols | Experimental Assay: koff ; Experimental Assay: kon ; Experimental Assay: KD |
Libraries | antibody data |
Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
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Structure ID | Release Date | Resolution | Structure Title |
---|---|---|---|
1BOG | 1998-08-04T00:00:00+0000 | 2.6 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-HOMOLOGOUS PEPTIDE |
1CFN | 1999-03-19T00:00:00+0000 | 2.65 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-RELATED PEPTIDE |
1CFQ | 1999-03-19T00:00:00+0000 | 2.8 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 |
1CFS | 1999-03-19T00:00:00+0000 | 2.75 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-UNRELATED PEPTIDE |
1CFT | 1999-03-19T00:00:00+0000 | 2.8 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-UNRELATED D-PEPTIDE |
1FRG | 1994-01-17T00:00:00+0000 | 2.8 | CRYSTAL STRUCTURE, SEQUENCE, AND EPITOPE MAPPING OF A PEPTIDE COMPLEX OF AN ANTI-INFLUENZA HA PEPTIDE ANTIBODY FAB 26(SLASH)9: FINE-TUNING ANTIBODY SPECIFICITY |
1HH6 | 2000-12-21T00:00:00+0000 | 2.6 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE |
1HH9 | 2000-12-21T00:00:00+0000 | 2.7 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE |
1HI6 | 2001-01-02T00:00:00+0000 | 2.55 | ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE |
1BFV | 1997-05-27T00:00:00+0000 | 2.1 | MONOCLONAL ANTIBODY FRAGMENT FV4155 FROM E. COLI |
Percent Identity | Matching Chains | Protein | Accession | Entry Name |
---|---|---|---|---|
95.5 | VL(gl) | P01631 | KV2A7_MOUSE | |
100.0 | H | Ig gamma-2A chain C region, membrane-bound form,4-4-20 (IG*G2A=KAPPA=) FAB FRAGMENT | P01863 | GCAA_MOUSE |
100.0 | H | Ig gamma-2A chain C region, membrane-bound form,4-4-20 (IG*G2A=KAPPA=) FAB FRAGMENT | P01865 | GCAM_MOUSE |
94.0 | H | Ig gamma-2A chain C region, membrane-bound form,4-4-20 (IG*G2A=KAPPA=) FAB FRAGMENT | P01864 | GCAB_MOUSE |
100.0 | L | Ig gamma-2A chain C region, membrane-bound form,4-4-20 (IG*G2A=KAPPA=) FAB FRAGMENT | P01837 | IGKC_MOUSE |