Natural and artificial mutants of the human 2,3-bisphosphoglycerate as a tool for the evaluation of structure-function relationships.
2,3-bisphosphoglycerate mutase is a multifunctional enzyme which catalyses in red blood cells the synthesis and the degradation of 2,3-bisphosphoglycerate, the allosteric effector of hemoglobin. In order to study the structure-function relationships in BPGM, an expression vector was constructed which yielded an active protein, but with a modified electrophoretic mobility, due to a non-blocked N-terminal residue. Using site directed mutagenesis, mutants were produced with shortened chains. Results indicated the importance of residues 252-256 for the function. A natural deficient mutant with the substitution 89 Arg----Cys was described. Artificial mutant with the same substitution reproduced the same defect, as well as mutants Arg----Gly and Arg----Ser, indicating the key role of Arg 89 in the enzymatic mechanism.
Garel
M C
MC
Unité INSERM-U91, Hôpital Henri Mondor, Créteil, France.
Lemarchandel
V
V
Prehu
M O
MO
Calvin
M C
MC
Arous
N
N
Rosa
R
R
Rosa
J
J
Cohen-Solal
M
M
D019794:2,3-Diphosphoglycerate; D000595:Amino Acid Sequence; D001483:Base Sequence; D001731:Bisphosphoglycerate Mutase / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004163:Diphosphoglyceric Acids / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010770:Phosphotransferases / Q000235:genetics*; D010957:Plasmids; D013329:Structure-Activity Relationship
D004163:Diphosphoglyceric Acids; D019794:2,3-Diphosphoglycerate; D004247:DNA; D010770:Phosphotransferases; D001731:Bisphosphoglycerate Mutase