Solution NMR structures of IgG binding domains with artificially evolved high levels of sequence identity but different folds

Biochemistry. 2005 Nov 1;44(43):14055-61. doi: 10.1021/bi051232j.

Abstract

We describe here the solution NMR structures of two IgG binding domains with highly homologous sequences but different three-dimensional structures. The proteins, G311 and A219, are derived from the IgG binding domains of their wild-type counterparts, protein G and protein A, respectively. Through a series of site-directed mutations and phage display selections, the sequences of G311 and A219 were designed to converge to a point of high-level sequence identity while keeping their respective wild-type tertiary folds. Structures of both artificially evolved sequences were determined by NMR spectroscopy. The main chain fold of G311 can be superimposed on the wild-type alpha/beta protein G structure with a backbone rmsd of 1.4 A, and the A219 structure can be overlaid on the wild-type three-alpha-helix protein A fold also with a backbone rmsd of 1.4 A. The structure of G311, in particular, accommodates a large number of mutational changes without undergoing a change in the overall fold of the main chain. The structural differences are maintained despite a high level (59%) of sequence identity. These proteins serve as starting points for further experiments that will probe basic concepts of protein folding and conformational switching.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Crystallography, X-Ray
  • Immunoglobulin G / chemistry*
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Mutation
  • Nerve Tissue Proteins / chemistry*
  • Protein Folding*
  • Protein Structure, Secondary
  • Staphylococcal Protein A / chemistry*
  • Thermodynamics

Substances

  • G-substrate
  • Immunoglobulin G
  • Nerve Tissue Proteins
  • Staphylococcal Protein A

Associated data

  • PDB/1ZXG
  • PDB/1ZXH