TY - JOUR
T1 - SERCA1 protein expression in muscle of patients with Brody disease and Brody syndrome and in cultured human muscle fibers
AU - Guglielmi, Valeria
AU - Vattemi, Gaetano
AU - Gualandi, Francesca
AU - Voermans, Nicol C.
AU - Marini, Matteo
AU - Scotton, Chiara
AU - Pegoraro, Elena
AU - Oosterhof, Arie
AU - Kósa, Magdolna
AU - Zádor, Erno
AU - Valente, Enza Maria
AU - De Grandis, Domenico
AU - Neri, Marcella
AU - Codemo, Valentina
AU - Novelli, Antonio
AU - van Kuppevelt, Toin H.
AU - Dallapiccola, Bruno
AU - van Engelen, Baziel G.
AU - Ferlini, Alessandra
AU - Tomelleri, Giuliano
PY - 2013/9
Y1 - 2013/9
N2 - Brody disease is an inherited myopathy associated with a defective function of sarcoplasmic/endoplasmic reticulum Ca2+-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.
AB - Brody disease is an inherited myopathy associated with a defective function of sarcoplasmic/endoplasmic reticulum Ca2+-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.
KW - ATP2A1 gene
KW - Brody disease
KW - Brody syndrome
KW - Sarcoplasmic/endoplasmic reticulum Ca-ATPase 1 (SERCA1)
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U2 - 10.1016/j.ymgme.2013.07.015
DO - 10.1016/j.ymgme.2013.07.015
M3 - Article
C2 - 23911890
AN - SCOPUS:84882868295
VL - 110
SP - 162
EP - 169
JO - Molecular Genetics and Metabolism
JF - Molecular Genetics and Metabolism
SN - 1096-7192
IS - 1-2
ER -