Hsp27-2D-gel electrophoresis is a diagnostic tool to differentiate primary desminopathies from myofibrillar myopathies.
Small heat shock proteins prevent abnormal protein folding and accumulation. We analyzed the expression of hsp27 and alphaB-crystallin in skeletal muscle specimens of patients with desminopathies, plectinopathies, myotilinopathy, and other myofibrillar myopathies by means of differential centrifugation, 2D-gel electrophoresis, Western blotting, and mass spectrometry. Hsp27-P82 and -P15 as well as alphaB-crystallin-P59 and -P45 are the major serine phosphorylation isoforms in normal and diseased human skeletal muscle. 2D-gel-electrophoresis revealed spots of hsp27 in a range of pH 5.3-6.4 in samples of all skeletal muscle specimens, except for the seven desminopathies. They indicated a shift of the main hsp27-spot to alkaline pH degrees, which may help to differentiate primary desminopathies from other myopathies with structural pathology of the desmin cytoskeleton.
Clemen
Christoph S
CS
Department of Neurology, Medical Faculty, University of Bonn, Sigmund Freud Str. 25, 53127 Bonn, FRG.
Fischer
Dirk
D
Roth
Udo
U
Simon
Stéphanie
S
Vicart
Patrick
P
Kato
Kanefusa
K
Kaminska
Anna M
AM
Vorgerd
Matthias
M
Goldfarb
Lev G
LG
Eymard
Bruno
B
Romero
Norma B
NB
Goudeau
Bertrand
B
Eggermann
Thomas
T
Zerres
Klaus
K
Noegel
Angelika A
AA
Schröder
Rolf
R
D000595:Amino Acid Sequence; D003893:Desmin / Q000032:analysis; D003937:Diagnosis, Differential; D015180:Electrophoresis, Gel, Two-Dimensional; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry / Q000473:pathology; D020914:Myopathies, Structural, Congenital / Q000175:diagnosis*; D009363:Neoplasm Proteins / Q000032:analysis* / Q000378:metabolism; D010766:Phosphorylation; D020033:Protein Isoforms / Q000032:analysis / Q000378:metabolism; D038203:alpha-Crystallin B Chain / Q000032:analysis / Q000378:metabolism
D003893:Desmin; D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D038203:alpha-Crystallin B Chain