Proteomic profiling of endothelin-1-stimulated hypertrophic cardiomyocytes reveals the increase of four different desmin species and alpha-B-crystallin.
We performed a proteomic investigation on primary cultures of neonatal rat cardiomyocytes after treatment with 10 nM endothelin-1 (ET1) for 48 h, an in vitro model for cardiac hypertrophy. Two-dimensional gel electrophoresis profiles of cell lysates were compared after colloidal Coomassie Blue staining. 12 protein spots that significantly changed in density due to ET1 stimulation were selected for in-gel digestion and identified through mass spectrometry. Of these, 8 spots were increased and 4 were decreased. Four of the increased proteins were identified as desmin, the cardiac component of intermediate filaments and one as alpha-B-crystallin, a molecular chaperone that binds desmin. All the desmins increased 2- to 5-fold, and alpha-B-crystallin increased 2-fold after ET1 treatment. Desmin cytoskeleton has been implicated in the regulation of mitochondrial activity and distribution, as well as in the formation of amyloid bodies. Mitochondria-specific fluorescent probe MitoTracker indicated mitochondrial redistribution in hypertrophic cells. An increase of amyloid aggregates containing desmin upon treatment with ET1 was detected by filter assay. Of the four proteins that showed decreased abundance after ET1 treatment, the chaperones hsp60 and grp75 were decreased 13- and 9-fold, respectively. In conclusion, proteomic profiling of ET1-stimulated rat neonatal cardiomyocytes reveals specific changes in cardiac molecular phenotype mainly involving intermediate filament and molecular chaperone proteins.
Agnetti
Giulio
G
Department of Biochemistry G. Moruzzi, INRC, University of Bologna, Italy. giulio.agnetti@unibo.it
Bezstarosti
Karel
K
Dekkers
Dick H W
DH
Verhoeven
Adrie J M
AJ
Giordano
Emanuele
E
Guarnieri
Carlo
C
Caldarera
Claudio M
CM
Van Eyk
Jennifer E
JE
Lamers
Jos M J
JM
D000818:Animals; D006332:Cardiomegaly / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D003459:Crystallins; D003893:Desmin / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D019332:Endothelin-1 / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology; D040901:Proteomics; D051381:Rats; D017208:Rats, Wistar; D021241:Spectrometry, Mass, Electrospray Ionization
D003459:Crystallins; D003893:Desmin; D019332:Endothelin-1; D006360:Heat-Shock Proteins; D008869:Microtubule-Associated Proteins; C530883:cryaB protein, rat