On the Potential Origins of the High Stability of Reconstructed Ancestral Proteins

Mol Biol Evol. 2016 Oct;33(10):2633-41. doi: 10.1093/molbev/msw138. Epub 2016 Jul 12.


Ancestral reconstruction provides instrumental insights regarding the biochemical and biophysical characteristics of past proteins. A striking observation relates to the remarkably high thermostability of reconstructed ancestors. The latter has been linked to high environmental temperatures in the Precambrian era, the era relating to most reconstructed proteins. We found that inferred ancestors of the serum paraoxonase (PON) enzyme family, including the mammalian ancestor, exhibit dramatically increased thermostabilities compared with the extant, human enzyme (up to 30 °C higher melting temperature). However, the environmental temperature at the time of emergence of mammals is presumed to be similar to the present one. Additionally, the mammalian PON ancestor has superior folding properties (kinetic stability)-unlike the extant mammalian PONs, it expresses in E. coli in a soluble and functional form, and at a high yield. We discuss two potential origins of this unexpectedly high stability. First, ancestral stability may be overestimated by a "consensus effect," whereby replacing amino acids that are rare in contemporary sequences with the amino acid most common in the family increases protein stability. Comparison to other reconstructed ancestors indicates that the consensus effect may bias some but not all reconstructions. Second, we note that high stability may relate to factors other than high environmental temperature such as oxidative stress or high radiation levels. Foremost, intrinsic factors such as high rates of genetic mutations and/or of transcriptional and translational errors, and less efficient protein quality control systems, may underlie the high kinetic and thermodynamic stability of past proteins.

Keywords: ancestral inference; consensus effect.; protein evolution; protein stability; serum paraoxonase (PON); thermostability.

MeSH terms

  • Amino Acid Sequence / genetics
  • Animals
  • Aryldialkylphosphatase / blood
  • Aryldialkylphosphatase / genetics*
  • Aryldialkylphosphatase / metabolism
  • Bacterial Proteins / genetics
  • Carboxylic Ester Hydrolases / genetics
  • Enzyme Stability
  • Escherichia coli / genetics
  • Evolution, Molecular
  • Hot Temperature
  • Humans
  • Phylogeny
  • Protein Stability
  • Sequence Alignment / methods


  • Bacterial Proteins
  • Carboxylic Ester Hydrolases
  • arylesterase
  • Aryldialkylphosphatase