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Eight novel mutations and consequences on mRNA and protein level in pyruvate kinase-deficient patients with nonspherocytic hemolytic anemia
Wilfried Kugler, Christian Willaschek,
Christiane Holtz, Andreas Ohlenbusch,
Petra Laspe, Regina Krügener, Hilary Muirhead,
Werner Schröter, Max Lakomek,
Christian Willaschek
Universitäts-Kinderklinik, Göttingen, Germany
Search for more papers by this authorAndreas Ohlenbusch
Universitäts-Kinderklinik, Göttingen, Germany
Search for more papers by this authorHilary Muirhead
Department of Biochemistry and Molecular Recognition Centre, University of Bristol, Bristol, UK
Search for more papers by this authorCorresponding Author
Max Lakomek
Universitäts-Kinderklinik, Göttingen, Germany
Universitäts-Kinderklinik, Robert-Koch-Str. 40, D-37075 Göttingen, GermanySearch for more papers by this authorWilfried Kugler, Christian Willaschek,
Christiane Holtz, Andreas Ohlenbusch,
Petra Laspe, Regina Krügener, Hilary Muirhead,
Werner Schröter, Max Lakomek,
Christian Willaschek
Universitäts-Kinderklinik, Göttingen, Germany
Search for more papers by this authorAndreas Ohlenbusch
Universitäts-Kinderklinik, Göttingen, Germany
Search for more papers by this authorHilary Muirhead
Department of Biochemistry and Molecular Recognition Centre, University of Bristol, Bristol, UK
Search for more papers by this authorCorresponding Author
Max Lakomek
Universitäts-Kinderklinik, Göttingen, Germany
Universitäts-Kinderklinik, Robert-Koch-Str. 40, D-37075 Göttingen, GermanySearch for more papers by this authorFirst published: 10 February 2000
Citations: 14
Abstract
Pyruvate kinase (PK) deficiency (PKD) is an autosomal recessive disorder with the typical manifestation of nonspherocytic hemolytic anemia. We analyzed the mutant enzymes of 10 unrelated patients with PKD, whose symptoms ranged from a mild, chronic hemolytic anemia to a severe anemia, by sequence analysis for the presence of alterations in the PKLR gene. In all cases the patients were shown to be compound heterozygous. Eight novel mutations were identified: 458T→C (Ile153Thr), 656T→C (Ile219Thr), 877G→A (Asp293Asn), 991G→A (Asp331Asn), 1055C→A (Ala352Asp), 1483G→A (Ala495Thr), 1649A→T (Asp550Val), and 183-184ins16bp. This 16 bp duplication produces a frameshift and subsequent stop codon resulting in a drastically reduced mRNA level, and probably in an unstable gene product. Surprisingly, the existence of M2-type PK could be demonstrated in the patient's red blood cells. The study of different polymorphic sites revealed, with one exception, a strict linkage of the 1705C, 1738T, IVS5(+51)T, T(10) polymorphisms and the presence of 14 ATT repeats in intron 11. Our analyses show the consequences of a distorted structure on enzyme function and we discuss the correlations between the mutations identified and the parameters indicative for enzyme function. Hum Mutat 15:261–272, 2000. © 2000 Wiley-Liss, Inc.
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