title: 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.affiliation: Department of Neurological, Neuropsychological, Morphological and Movement Sciences, Section of Clinical Neurology, University of Verona, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Guglielmi
authors.forename: Gaetano
authors.initials: G
authors.lastname: Vattemi
authors.forename: Francesca
authors.initials: F
authors.lastname: Gualandi
authors.forename: Nicol C
authors.initials: NC
authors.lastname: Voermans
authors.forename: Matteo
authors.initials: M
authors.lastname: Marini
authors.forename: Chiara
authors.initials: C
authors.lastname: Scotton
authors.forename: Elena
authors.initials: E
authors.lastname: Pegoraro
authors.forename: Arie
authors.initials: A
authors.lastname: Oosterhof
authors.forename: Magdolna
authors.initials: M
authors.lastname: Kósa
authors.forename: Ernő
authors.initials: E
authors.lastname: Zádor
authors.forename: Enza Maria
authors.initials: EM
authors.lastname: Valente
authors.forename: Domenico
authors.initials: D
authors.lastname: De Grandis
authors.forename: Marcella
authors.initials: M
authors.lastname: Neri
authors.forename: Valentina
authors.initials: V
authors.lastname: Codemo
authors.forename: Antonio
authors.initials: A
authors.lastname: Novelli
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallapiccola
authors.forename: Baziel G
authors.initials: BG
authors.lastname: van Engelen
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ferlini
authors.forename: Giuliano
authors.initials: G
authors.lastname: Tomelleri
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D002478:Cells, Cultured; D002675:Child, Preschool; D005091:Exons; D005260:Female; D005786:Gene Expression Regulation; D005838:Genotype; D006801:Humans; D007223:Infant; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000473:pathology; D009154:Mutation; D009224:Myotonia Congenita / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000235:genetics*; D046509:Tissue Culture Techniques
chemical_list: D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; C506943:ATP2A1 protein, human
keywords: ATP2A1 gene; Brody disease; Brody syndrome; Sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase 1 (SERCA1)