title: The contribution of coevolving residues to the stability of KDO8P synthase.
abstract: BACKGROUND
  The evolutionary tree of 3-deoxy-D-manno-octulosonate 8-phosphate (KDO8P) synthase (KDO8PS), a bacterial enzyme that catalyzes a key step in the biosynthesis of bacterial endotoxin, is evenly divided between metal and non-metal forms, both having similar structures, but diverging in various degrees in amino acid sequence. Mutagenesis, crystallographic and computational studies have established that only a few residues determine whether or not KDO8PS requires a metal for function. The remaining divergence in the amino acid sequence of KDO8PSs is apparently unrelated to the underlying catalytic mechanism.
  
  
  RESULTS
  The multiple alignment of all known KDO8PS sequences reveals that several residue pairs coevolved, an indication of their possible linkage to a structural constraint. In this study we investigated by computational means the contribution of coevolving residues to the stability of KDO8PS. We found that about 1/4 of all strongly coevolving pairs probably originated from cycles of mutation (decreasing stability) and suppression (restoring it), while the remaining pairs are best explained by a succession of neutral or nearly neutral covarions.
  
  
  CONCLUSIONS
  Both sequence conservation and coevolution are involved in the preservation of the core structure of KDO8PS, but the contribution of coevolving residues is, in proportion, smaller. This is because small stability gains or losses associated with selection of certain residues in some regions of the stability landscape of KDO8PS are easily offset by a large number of possible changes in other regions. While this effect increases the tolerance of KDO8PS to deleterious mutations, it also decreases the probability that specific pairs of residues could have a strong contribution to the thermodynamic stability of the protein.
authors.affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan, United States of America.
authors.forename: Sharon H
authors.initials: SH
authors.lastname: Ackerman
authors.forename: Domenico L
authors.initials: DL
authors.lastname: Gatti
mesh_terms: D000446:Aldehyde-Lyases / Q000235:genetics* / Q000378:metabolism*; D000596:Amino Acids / Q000235:genetics*; D001419:Bacteria / Q000201:enzymology; D004795:Enzyme Stability; D019143:Evolution, Molecular; D008390:Markov Chains; D008670:Metals / Q000378:metabolism
chemical_list: D000596:Amino Acids; D008670:Metals; C020011:2-dehydro-3-deoxyphosphooctonate aldolase; D000446:Aldehyde-Lyases