title: Red cell pyruvate kinase deficiency: 17 new mutations of the PK-LR gene.
abstract: The PK-LR gene was studied in 23 patients with congenital haemolytic anaemia associated with erythrocyte pyruvate kinase (PK) deficiency. Twenty-seven different mutations were detected among the 42 mutated alleles identified: 19 missense mutations, four splice site mutations and one nonsense, one single base deletion and two large deletions. Seventeen of them (107G, 278T, 403T, 409A, 661A, 859C, 958A, 1094T, 1190T, 1209A, 1232C, 1369G, 507A, IVS9 -1c, IVS9 +43c [corrected] del C224, del 5006bp IVS3--> nt 1431) were new. Although all the exons, the flanking regions and the promoter were sequenced in all cases, we failed to detect the second expected mutation in four subjects. To correlate genotype to phenotype, the molecular results were related to the biochemical properties of the mutant enzymes by an analysis of the three-dimensional structure of erythrocyte PK. The new mutant 409A, found in association with the large deletion of 5006 bp in a newborn baby who died soon after birth, was functionally characterized by mutagenesis and in vitro expression of the protein to investigate its contribution in the severity of the clinical pattern. However, the biochemical data obtained for the mutant enzyme cannot explain the severe anaemia found in the PK-deficient patient hemizygous for this mutation.
authors.affiliation: Dipartimento di Ematologia, IRCCS Ospedale Maggiore di Milano, Milan, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Fermo
authors.forename: Paola
authors.initials: P
authors.lastname: Bianchi
authors.forename: Laurent R
authors.initials: LR
authors.lastname: Chiarelli
authors.forename: Frederic
authors.initials: F
authors.lastname: Cotton
authors.forename: Cristina
authors.initials: C
authors.lastname: Vercellati
authors.forename: Karin
authors.initials: K
authors.lastname: Writzl
authors.forename: Kerry
authors.initials: K
authors.lastname: Baker
authors.forename: Ian
authors.initials: I
authors.lastname: Hann
authors.forename: Robin
authors.initials: R
authors.lastname: Rodwell
authors.forename: Giovanna
authors.initials: G
authors.lastname: Valentini
authors.forename: Alberto
authors.initials: A
authors.lastname: Zanella
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000745:Anemia, Hemolytic, Congenital / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis; D004912:Erythrocytes / Q000201:enzymology*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D010375:Pedigree; D011770:Pyruvate Kinase / Q000737:chemistry / Q000172:deficiency* / Q000235:genetics*
chemical_list: D011770:Pyruvate Kinase