Compound heterozygosity in a complete erythrocyte bisphosphoglycerate mutase deficiency

Abstract

Erythrocyte bisphosphoglycerate mutase (BPGM) deficiency is a rare disease associated with a decrease in 2,3-diphosphoglycerate concentration. A complete BPGM deficiency was described in 1978 by Rosa et al (J Clin Invest 62:907, 1978) and was shown to be associated with 30% to 50% of an inactive enzyme detectable by specific antibodies and resulting from an 89 Arg-->Cys substitution. The propositus' three sisters exhibited the same phenotype, while his two children had an intermediate phenotype. Samples from the family were examined using polymerase chain reaction and allele-specific oligonucleotide hybridization and sequencing techniques. Amplification of erythrocyte total RNA from the propositus' sister around the 89 mutation indicated the presence of two forms of messenger RNAs, a major form with the 89 Arg-->Cys mutation and a minor form with a normal sequence. Sequence studies of the propositus' DNA samples indicated heterozygosity at locus 89 and another heterozygosity with the deletion of nucleotide C 205 or C 206. Therefore, the total BPGM deficiency results from a genetic compound with one allele coding for an inactive enzyme (mutation BPGM Creteil I) and the other bearing a frameshift mutation (mutation BPGM Creteil II). Examination of the propositus' two children indicated that they both inherited the BPGM Creteil I mutation.

Erythrocytosis due to bisphosphoglycerate mutase deficiency with concurrent glucose‐6‐phosphate dehydrogenase (G‐6‐PD) deficiency

“…The first mutation was an arginine to cysteine mutation at amino acid 89 (corresponding to a C!T substitution at nucleotide 413); the second mutation was a frame shift mutation. Analysis of the propositus' children showed only the presence of the 89 Arg!Cys mutation [5]. The parents of the propositus were deceased and thus could not be evaluated.…”

“…The three exons of the BPGM gene were amplified from genomic DNA obtained from peripheral blood leukocytes using the polymerase chain reaction (PCR). Exons 1 and 2 were amplified using the primers described by Lemarchandel et al [5], with an annealing temperature of 56 C for exon 1 and of 52 C for exon 2. Exon 3 was amplified using 5 0 -TGATGTAGCA-CTTGCTGTG-3 0 and 5 0 -GTGAACTACTGATTAG-AATAGTG-3 0 , with a 55 C annealing temperature, giving a 762-bp fragment.…”

Five unknown mutations in the LR pyruvate kinase gene associated with severe hereditary nonspherocytic haemolytic anaemia in France

“…Fresh blood was collected in tubes containing EDTA. DNA was extracted from leucocytes and prepared using standard techniques (Lemarchandel et al, 1992).…”

“…The coding regions, intron-exon boundaries, promoter and small introns, of the PK gene were amplified using four sets of oligonucleotides for 30 cycles (948C for 1 min, 55-678C for 1 min and 728C for 1 min 30 s with 1-2 mM MgCl 2 ) ( Fig 1, Table I). Single-stranded DNA was obtained by an asymmetric PCR reaction using similar conditions (60 cycles with a primer ratio of 50 : 2) (Lemarchandel et al, 1992).…”

Molecular basis of erythroenzymopathies associated with hereditary hemolytic anemia: Tabulation of mutant enzymes

“…A complete deficiency of DPGM was discovered in a man of French origin associated with a moderate erythrocytosis. Sequence studies of this individual indicated heterozygosity with a cytidine to thymine substitution at nucleotide 413 and another heterozygosity with the deletion of cytidine at nucleotide 205 or 206 [93]. Therefore, the complete enzyme deficiency resulted from a genetic compound with one allele coding for a missense mutation (Arg to Cys at 89) and the other bearing a frameshift and premature termination of translation.…”