Focused proteomics revealed a novel Rho-kinase signaling pathway in the heart

Yoshimitsu Yura, Mutsuki Amano, Mikito Takefuji, Tomohiro Bando, Kou Suzuki, Katsuhiro Kato, Tomonari Hamaguchi, Md Hasanuzzaman Shohag, Tetsuya Takano, Yasuhiro Funahashi, Shinichi Nakamuta, Keisuke Kuroda, Tomoki Nishioka, Toyoaki Murohara, Kozo Kaibuchi

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)105-120
Number of pages16
JournalCell structure and function
Issue number2
Publication statusPublished - 2016
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Physiology
  • Molecular Biology
  • Cell Biology


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