Mutations in Czech LGMD2A patients revealed by analysis of calpain3 mRNA and their phenotypic outcome.
Calpain3 (CAPN3, p94) is a muscle-specific nonlysosomal cysteine proteinase. Loss of proteolytic function or change of other properties of this enzyme (such as stability or ability to interact with other muscular proteins) is manifested as limb girdle muscular dystrophy type 2A (LGMD2A, calpainopathy). These pathological changes in properties of calpain3 are caused by mutations in the calpain3 gene. The fact that the human gene for calpain3 is quite long led us to analyse its coding sequence by reverse transcription-PCR followed by sequence analysis. This study reports nine mutations that we found by analysing mRNA of seven unrelated LGMD patients in the Czech Republic. Three of these mutations were novel, not described on the Leiden muscular dystrophy pages so far. Further, we observed a reduction of dysferlin in muscle membrane in five of our seven LGMD2A patients by immunohistochemical analysis of muscle sections.
Chrobáková
Tána
T
Center of Molecular Biology and Gene Therapy, University Hospital Brno, Cernopolni 9, CZ-62500 Brno, Czech Republic.
Hermanová
Markéta
M
Kroupová
Iva
I
Vondrácek
Petr
P
Maríková
Tat'ána
T
Mazanec
Radim
R
Zámecník
Josef
J
Stanek
Jan
J
Havlová
Miluse
M
Fajkusová
Lenka
L
D000293:Adolescent; D000328:Adult; D000368:Aged; D001120:Arginine / Q000235:genetics; D015153:Blotting, Western / Q000379:methods; D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D002648:Child; D018153:Czech Republic; D004252:DNA Mutational Analysis / Q000379:methods; D000073939:Dysferlin; D005091:Exons / Q000235:genetics; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008875:Middle Aged; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014364:Tryptophan / Q000235:genetics
C114839:DYSF protein, human; D000073939:Dysferlin; D007527:Isoenzymes; D008565:Membrane Proteins; D009124:Muscle Proteins; D012333:RNA, Messenger; D014364:Tryptophan; D001120:Arginine; C105884:CAPN3 protein, human; D002154:Calpain