title: Molecular abnormalities of a phosphoglycerate kinase variant generated by spontaneous mutation.
abstract: A new case of X chromosome-linked phosphoglycerate kinase (PGK) abnormality is described. The male proband was mentally retarded, had behavior disorders, and displayed episodes of hemolytic anemia. The enzyme activity of red blood cells from the patient was about 10% of normal, and that of the cultured fibroblasts was about 50% of normal cells. The variant PGK was characterized by a lower affinity for the substrates, reduced thermostability, and increased anodal electrophoretic mobility. The pH activity profile of the variant enzyme was different from that of normal. The amount of messenger RNA (mRNA) in the variant fibroblasts was comparable to that of normal fibroblasts. The mRNA coding for PGK was subjected to coupled reverse transcription followed by amplification by the polymerase chain reaction. Nucleotide sequence of the variant cDNA showed a point mutation, T/A----C/G transition, in exon 9 of the variant gene. No other mutation was found in all coding regions of the variant. The mutation should cause Cys----Arg substitution at the 315th position from the NH2-terminal Ser of PGK, and it created an additional Ava II (or isoschimatic) cleavage site in the variant gene. Because the variant gene was not detected in the proband's mother and siblings, it must have been generated by spontaneous mutation during oogenesis.
authors.affiliation: Department of Biochemical Genetics, Beckman Research Institute, City of Hope, Duarte, CA 91010.
authors.forename: M
authors.initials: M
authors.lastname: Maeda
authors.forename: E V
authors.initials: EV
authors.lastname: Bawle
authors.forename: R
authors.initials: R
authors.lastname: Kulkarni
authors.forename: E
authors.initials: E
authors.lastname: Beutler
authors.forename: A
authors.initials: A
authors.lastname: Yoshida
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D002869:Chromosome Aberrations; D025063:Chromosome Disorders; D004247:DNA / Q000235:genetics; D005347:Fibroblasts / Q000201:enzymology; D014644:Genetic Variation; D015698:Genomic Library; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D010735:Phosphoglycerate Kinase / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012016:Reference Values; D015183:Restriction Mapping; D014960:X Chromosome
chemical_list: D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D004247:DNA; D005954:Glucosephosphate Dehydrogenase; D010735:Phosphoglycerate Kinase