| Abstract | Limb-girdle muscular dystrophy type 2A (LGMD2A) is an autosomal recessive muscular disorder caused by mutations in the gene coding for calpain 3, a calcium-dependent protease. We developed an in vitro assay that can detect the proteolytic activity of calpain 3 in a muscle sample. This assay is based on the use of an inactive calpain 3 as a substrate for active calpain 3 molecules. A total of 79 human biopsies have been analysed using an unbiased single blind method. Results were confronted with the molecular diagnosis for confirmation. Proteolytic activity was either reduced or absent in 68% of LGMD2A biopsies. In the remaining 32%, normal proteolytic activity was found despite the presence of calpain 3 mutation(s), suggesting that other calpain 3 properties might be impaired to give rise to the LGMD2A phenotype. Our assay is easily adaptable to routine and appears to be more sensitive than common analysis by immunodetection. |
| Authors | Astrid Milic, Nathalie Daniele, Hanns Lochmüller, Marina Mora, Giacomo P Comi, Maurizio Moggio, Fanny Noulet, Maggie C Walter, Lucia Morandi, Jérôme Poupiot, Carinne Roudaut, Reginald E Bittner, Marc Bartoli, Isabelle Richard
(Affiliation: Laboratory of Clinical Neurogenetics and Muscular Disorders, Zagreb University School of Medicine, Croatian Institute for Brain Research, Salata 12, 10000 Zagreb, Croatia.)
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| Journal | Neuromuscular disorders : NMD
(Neuromuscul Disord)
Vol. 17
Issue 2
Pg. 148-56
(Feb 2007)
ISSN: 0960-8966 England |
| PMID | 17236769
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
| Chemical References |
- Muscle Proteins
- CAPN3 protein, human
- Calpain
|
| Topics |
- Animals
- Blotting, Western
- Calpain
(metabolism)
- Disease Models, Animal
- Electrophoresis, Polyacrylamide Gel
- Heterozygote
- Humans
- Mice
- Muscle Proteins
(metabolism)
- Muscle, Skeletal
(enzymology)
- Muscular Dystrophies, Limb-Girdle
(enzymology, genetics, pathology)
- NIH 3T3 Cells
- Phenotype
- Reproducibility of Results
- Tissue Banks
- Transfection
|
