TY - JOUR
T1 - Proteomic identification of FHL1 as the protein mutated in human reducing body myopathy
AU - Schessl, Joachim
AU - Zou, Yaqun
AU - Mcgrath, Meagan Jane
AU - Cowling, Belinda Simone
AU - Maiti, Baijayanta
AU - Chin, Steven S
AU - Sewry, Caroline
AU - Battini, Roberta
AU - Hu, Ying
AU - Cottle, Denny
AU - Rosenblatt, Michael
AU - Spruce, Lynn
AU - Ganguly, Arupa
AU - Kirschner, Janbernd
AU - Judkins, Alexander R
AU - Golden, Jeffrey A
AU - Goebel, Hans-Hilmar
AU - Muntoni, Francesco
AU - Flanigan, Kevin M
AU - Mitchell, Christina Anne
AU - Bonnemann, Carsten G
PY - 2008
Y1 - 2008
N2 - Reducing body myopathy (RBM) is a rare disorder causing progressive muscular weakness characterized by aggresome-like inclusions in the myofibrils. Identification of genes responsible for RBM by traditional genetic approaches has been impossible due to the frequently sporadic occurrence in affected patients and small family sizes. As an alternative approach to gene identification, we used laser microdissection of intracytoplasmic inclusions identified in patient muscle biopsies, followed by nanoflow liquid chromatography-tandem mass spectrometry and proteomic analysis. The most prominent component of the inclusions was the Xq26.3-encoded four and a half LIM domain 1 (FHL1) protein, expressed predominantly in skeletal but also in cardiac muscle. Mutational analysis identified 4 FHL1 mutations in 2 sporadic unrelated females and in 2 families with severely affected boys and less-affected mothers. Transfection of kidney COS-7 and skeletal muscle C2C12 cells with mutant FHL1 induced the formation of aggresome-like inclusions that incorporated both mutant and wild-type FHL1 and trapped other proteins in a dominant-negative manner. Thus, a novel laser microdissection/proteomics approach has helped identify both inherited and de novo mutations in FHL1, thereby defining a new X-linked protein aggregation disorder of muscle.
AB - Reducing body myopathy (RBM) is a rare disorder causing progressive muscular weakness characterized by aggresome-like inclusions in the myofibrils. Identification of genes responsible for RBM by traditional genetic approaches has been impossible due to the frequently sporadic occurrence in affected patients and small family sizes. As an alternative approach to gene identification, we used laser microdissection of intracytoplasmic inclusions identified in patient muscle biopsies, followed by nanoflow liquid chromatography-tandem mass spectrometry and proteomic analysis. The most prominent component of the inclusions was the Xq26.3-encoded four and a half LIM domain 1 (FHL1) protein, expressed predominantly in skeletal but also in cardiac muscle. Mutational analysis identified 4 FHL1 mutations in 2 sporadic unrelated females and in 2 families with severely affected boys and less-affected mothers. Transfection of kidney COS-7 and skeletal muscle C2C12 cells with mutant FHL1 induced the formation of aggresome-like inclusions that incorporated both mutant and wild-type FHL1 and trapped other proteins in a dominant-negative manner. Thus, a novel laser microdissection/proteomics approach has helped identify both inherited and de novo mutations in FHL1, thereby defining a new X-linked protein aggregation disorder of muscle.
UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18274675
U2 - 10.1172/JCI34450
DO - 10.1172/JCI34450
M3 - Article
SN - 0021-9738
VL - 118
SP - 904
EP - 912
JO - The Journal of Clinical Investigation
JF - The Journal of Clinical Investigation
IS - 3
ER -