SERCA1 protein expression in muscle of patients with Brody disease and Brody syndrome and in cultured human muscle fibers.

Abstract

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.

• Authors: Valeria Guglielmi, Gaetano Vattemi, Francesca Gualandi, Nicol C Voermans, Matteo Marini, Chiara Scotton, Elena Pegoraro, Arie Oosterhof, Magdolna Kósa, Ernő Zádor, Enza Maria Valente, Domenico De Grandis, Marcella Neri, Valentina Codemo, Antonio Novelli, Toin H van Kuppevelt, Bruno Dallapiccola, Baziel G van Engelen, Alessandra Ferlini, Giuliano Tomelleri
• Journal: Molecular genetics and metabolism (Mol Genet Metab) 2013 Sep-Oct Vol. 110 Issue 1-2 Pg. 162-9 ISSN: 1096-7206 [Electronic] United States
• PMID: 23911890 (Publication Type: Journal Article)
• Copyright: © 2013 Elsevier Inc. All rights reserved.

Chemical References

• Sarcoplasmic Reticulum Calcium-Transporting ATPases
• ATP2A1 protein, human

Topics

• Adult
• Amino Acid Sequence
• Cells, Cultured
• Child, Preschool
• Exons
• Female
• Gene Expression Regulation
• Genotype
• Humans
• Infant
• Male
• Muscle Fibers, Skeletal (metabolism, pathology)
• Mutation
• Myotonia Congenita (diagnosis, genetics, pathology)
• Sarcoplasmic Reticulum Calcium-Transporting ATPases (genetics)
• Tissue Culture Techniques