1. SERCA1 protein expression in muscle of patients with Brody disease and Brody syndrome and in cultured human muscle fibers
- Author
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Nicol C. Voermans, Valeria Guglielmi, Elena Pegoraro, Antonio Novelli, C. Scotton, Domenico De Grandis, Bruno Dallapiccola, Alessandra Ferlini, Ernő Zádor, Marcella Neri, Arie Oosterhof, Gaetano Vattemi, Valentina Codemo, Francesca Gualandi, Baziel G.M. van Engelen, Giuliano Tomelleri, Magdolna Kósa, Toin H. van Kuppevelt, Enza Maria Valente, and Matteo Marini
- Subjects
Protein isoform ,Adult ,Male ,medicine.medical_specialty ,ATP2A1 gene ,Genotype ,Myotonia Congenita ,Endocrinology, Diabetes and Metabolism ,DCN MP - Plasticity and memory ,Sarcoplasm ,Muscle Fibers, Skeletal ,Biology ,Biochemistry ,Sarcoplasmic Reticulum Calcium-Transporting ATPases ,Tissue Culture Techniques ,Exon ,Endocrinology ,Internal medicine ,Genetics ,medicine ,Brody syndrome ,Humans ,Amino Acid Sequence ,Myopathy ,Molecular Biology ,Gene ,Brody disease ,Cells, Cultured ,Endoplasmic reticulum ,Alternative splicing ,Wild type ,Sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase 1 (SERCA1) ,SERCA1 ,Infant ,Exons ,Tissue engineering and pathology [NCMLS 3] ,Human Movement & Fatigue [DCN MP - Plasticity and memory NCEBP 10] ,Gene Expression Regulation ,Child, Preschool ,Mutation ,Female ,medicine.symptom - Abstract
Item does not contain fulltext Brody disease is an inherited myopathy associated with a defective function of sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase 1 (SERCA1) protein. Mutations in the ATP2A1 gene have been reported only in some patients. Therefore it has been proposed to distinguish patients with ATP2A1 mutations, Brody disease (BD), from patients without mutations, Brody syndrome (BS). We performed a detailed study of SERCA1 protein expression in muscle of patients with BD and BS, and evaluated the alternative splicing of SERCA1 in primary cultures of normal human muscle and in infant muscle. SERCA1 reactivity was observed in type 2 muscle fibers of patients with and without ATP2A1 mutations and staining intensity was similar in patients and controls. Immunoblot analysis showed a significant reduction of SERCA1 band in muscle of BD patients. In addition we demonstrated that the wild type and mutated protein exhibits similar solubility properties and that RIPA buffer improves the recovery of the wild type and mutated SERCA1 protein. We found that SERCA1b, the SERCA1 neonatal form, is the main protein isoform expressed in cultured human muscle fibers and infant muscle. Finally, we identified two novel heterozygous mutations within exon 3 of the ATP2A1 gene from a previously described patient with BD.
- Published
- 2013