TY - JOUR
T1 - Focused proteomics revealed a novel Rho-kinase signaling pathway in the heart
AU - Yura, Yoshimitsu
AU - Amano, Mutsuki
AU - Takefuji, Mikito
AU - Bando, Tomohiro
AU - Suzuki, Kou
AU - Kato, Katsuhiro
AU - Hamaguchi, Tomonari
AU - Shohag, Md Hasanuzzaman
AU - Takano, Tetsuya
AU - Funahashi, Yasuhiro
AU - Nakamuta, Shinichi
AU - Kuroda, Keisuke
AU - Nishioka, Tomoki
AU - Murohara, Toyoaki
AU - Kaibuchi, Kozo
N1 - Publisher Copyright:
© 2016 by Japan Society for Cell Biology.
PY - 2016
Y1 - 2016
N2 - Protein phosphorylation plays an important role in the physiological regulation of cardiac function. Myocardial contraction and pathogenesis of cardiac diseases have been reported to be associated with adaptive or maladaptive protein phosphorylation; however, phosphorylation signaling in the heart is not fully elucidated. We recently developed a novel kinase-interacting substrate screening (KISS) method for exhaustive screening of protein kinase substrates, using mass spectrometry and affinity chromatography. First, we examined protein phosphorylation by extracellular signal-regulated kinase (ERK) and protein kinase A (PKA), which has been relatively well studied in cardiomyocytes. The KISS method showed that ERK and PKA mediated the phosphorylation of known cardiac-substrates of each kinase such as Rps6ka1 and cTnI, respectively. Using this method, we found about 330 proteins as Rho-kinase-mediated substrates, whose substrate in cardiomyocytes is unknown. Among them, CARP/Ankrd1, a muscle ankyrin repeat protein, was confirmed as a novel Rho-kinasemediated substrate. We also found that non-phosphorylatable form of CARP repressed cardiac hypertrophyrelated gene Myosin light chain-2v (MLC-2v) promoter activity, and decreased cell size of heart derived H9c2 myoblasts more efficiently than wild type-CARP. Thus, focused proteomics enable us to reveal a novel signaling pathway in the heart.
AB - Protein phosphorylation plays an important role in the physiological regulation of cardiac function. Myocardial contraction and pathogenesis of cardiac diseases have been reported to be associated with adaptive or maladaptive protein phosphorylation; however, phosphorylation signaling in the heart is not fully elucidated. We recently developed a novel kinase-interacting substrate screening (KISS) method for exhaustive screening of protein kinase substrates, using mass spectrometry and affinity chromatography. First, we examined protein phosphorylation by extracellular signal-regulated kinase (ERK) and protein kinase A (PKA), which has been relatively well studied in cardiomyocytes. The KISS method showed that ERK and PKA mediated the phosphorylation of known cardiac-substrates of each kinase such as Rps6ka1 and cTnI, respectively. Using this method, we found about 330 proteins as Rho-kinase-mediated substrates, whose substrate in cardiomyocytes is unknown. Among them, CARP/Ankrd1, a muscle ankyrin repeat protein, was confirmed as a novel Rho-kinasemediated substrate. We also found that non-phosphorylatable form of CARP repressed cardiac hypertrophyrelated gene Myosin light chain-2v (MLC-2v) promoter activity, and decreased cell size of heart derived H9c2 myoblasts more efficiently than wild type-CARP. Thus, focused proteomics enable us to reveal a novel signaling pathway in the heart.
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U2 - 10.1247/csf.16011
DO - 10.1247/csf.16011
M3 - Article
C2 - 27334702
AN - SCOPUS:84984853848
SN - 0386-7196
VL - 41
SP - 105
EP - 120
JO - Cell structure and function
JF - Cell structure and function
IS - 2
ER -