Mining for allosteric information: natural mutations and positional sequence conservation in pyruvate kinase.
Although the amino acid sequences and the structures of pyruvate kinase (PYK) isozymes are highly conserved, allosteric regulations differ. This suggests that amino acids with low conservation play important roles in the allosteric mechanism. The current work exploits a 'natural screen'- the 122 point mutations identified in the human gene encoding the erythrocyte PYK isozyme and associated with nonspherocytic hemolytic anemia - to learn what amino acid positions in PYK may be important for allosteric regulations. In addition to the mutations, we consider the conservation of each amino acid position across 241 PYK sequences. Three groups of residue positions have been created, those with: (1) no disease causing mutation identified; (2) a disease causing mutation identified and high conservation across isozymes; and (3) a disease causing mutation identified and low conservation. Mutations at positions not identified in the natural screen are likely to be tolerated with minimal loss of function. Mutations at highly conserved positions are more likely to disrupt properties common to all PYK isozymes (e.g., structure, catalysis). Residues in the third group are likely to be involved in roles that are necessary for function but not common to all isozymes (e.g., allostery). Many of the Group 3 residues are located in the C-domain and to a lesser extent the A domain.
Department of Biochemistry and Molecular Biology, The University of Kansas Medical Center, Kansas City, USA.
David C
DC
Pendergrass
Rachel
R
Williams
James B
JB
Blair
Aron W
AW
Fenton
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000746:Anemia, Hemolytic, Congenital Nonspherocytic / Q000235:genetics*; D017124:Conserved Sequence / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D011770:Pyruvate Kinase / Q000737:chemistry* / Q000235:genetics*
D011770:Pyruvate Kinase