Enhancing Human Spermine Synthase Activity by Engineered Mutations

PLoS Comput Biol. 2013;9(2):e1002924. doi: 10.1371/journal.pcbi.1002924. Epub 2013 Feb 28.

Abstract

Spermine synthase (SMS) is an enzyme which function is to convert spermidine into spermine. It was shown that gene defects resulting in amino acid changes of the wild type SMS cause Snyder-Robinson syndrome, which is a mild-to-moderate mental disability associated with osteoporosis, facial asymmetry, thin habitus, hypotonia, and a nonspecific movement disorder. These disease-causing missense mutations were demonstrated, both in silico and in vitro, to affect the wild type function of SMS by either destabilizing the SMS dimer/monomer or directly affecting the hydrogen bond network of the active site of SMS. In contrast to these studies, here we report an artificial engineering of a more efficient SMS variant by transferring sequence information from another organism. It is confirmed experimentally that the variant, bearing four amino acid substitutions, is catalytically more active than the wild type. The increased functionality is attributed to enhanced monomer stability, lowering the pKa of proton donor catalytic residue, optimized spatial distribution of the electrostatic potential around the SMS with respect to substrates, and increase of the frequency of mechanical vibration of the clefts presumed to be the gates toward the active sites. The study demonstrates that wild type SMS is not particularly evolutionarily optimized with respect to the reaction spermidine → spermine. Having in mind that currently there are no variations (non-synonymous single nucleotide polymorphism, nsSNP) detected in healthy individuals, it can be speculated that the human SMS function is precisely tuned toward its wild type and any deviation is unwanted and disease-causing.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Computer Simulation
  • Humans
  • Hydrogen Bonding
  • Mental Retardation, X-Linked
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Stability
  • Sequence Alignment
  • Spermine Synthase / chemistry
  • Spermine Synthase / genetics*
  • Spermine Synthase / metabolism*
  • Static Electricity

Substances

  • Spermine Synthase

Supplementary concepts

  • Snyder Robinson syndrome