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Integrated Quantitative Phosphoproteomics and Cell-Based Functional Screening Reveals Specific Pathological Cardiac Hypertrophy-Related Phosphorylation Sites

  • Kwon, Hye Kyeong;Choi, Hyunwoo;Park, Sung-Gyoo;Park, Woo Jin;Kim, Do Han;Park, Zee-Yong
    • Molecules and Cells
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    • v.44 no.7
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    • pp.500-516
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    • 2021

  • Cardiac hypertrophic signaling cascades resulting in heart failure diseases are mediated by protein phosphorylation. Recent developments in mass spectrometry-based phosphoproteomics have led to the identification of thousands of differentially phosphorylated proteins and their phosphorylation sites. However, functional studies of these differentially phosphorylated proteins have not been conducted in a large-scale or high-throughput manner due to a lack of methods capable of revealing the functional relevance of each phosphorylation site. In this study, an integrated approach combining quantitative phosphoproteomics and cell-based functional screening using phosphorylation competition peptides was developed. A pathological cardiac hypertrophy model, junctate-1 transgenic mice and control mice, were analyzed using label-free quantitative phosphoproteomics to identify differentially phosphorylated proteins and sites. A cell-based functional assay system measuring hypertrophic cell growth of neonatal rat ventricle cardiomyocytes (NRVMs) following phenylephrine treatment was applied, and changes in phosphorylation of individual differentially phosphorylated sites were induced by incorporation of phosphorylation competition peptides conjugated with cell-penetrating peptides. Cell-based functional screening against 18 selected phosphorylation sites identified three phosphorylation sites (Ser-98, Ser-179 of Ldb3, and Ser-1146 of palladin) displaying near-complete inhibition of cardiac hypertrophic growth of NRVMs. Changes in phosphorylation levels of Ser-98 and Ser-179 in Ldb3 were further confirmed in NRVMs and other pathological/physiological hypertrophy models, including transverse aortic constriction and swimming models, using site-specific phospho-antibodies. Our integrated approach can be used to identify functionally important phosphorylation sites among differentially phosphorylated sites, and unlike conventional approaches, it is easily applicable for large-scale and/or high-throughput analyses.

  • https://doi.org/10.14348/molcells.2021.4002 인용 PDF KSCI

Appropriate Technology and Field Application of Non-powered Water Purification System Using Nanofiber Membrane (나노섬유 멤브레인 기반 무동력 정수 시스템의 적정기술 및 현장 적용)

  • Lee, Jin;Yun, Byeong Gweon;Han, Kyoung Gu;Lee, Seung Hoon;Kim, Cheol Hyeon;Kim, Chan;Lee, Yunho;Lee, Dongwhi;Lee, Seunghyeok;Kim, Kyoung-Woong
    • Journal of Appropriate Technology
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    • v.7 no.1
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    • pp.72-81
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    • 2021

  • Gravity-driven membrane (GDM) filtration system based on the nanofiber membrane was investigated. This system can be operated with little energy demand due to a gravitational pressure-driven filtration and biological fouling control strategy. Moreover, the optimal module configuration based on the high permeance of nanofiber membrane can provide a significantly high water productivity. In order to evaluate its applicability potential, the pilot-scale (3000-5000 L/day) systems with nanofiber membranes were operated in developing countries (Kiribati and Tuvalu). Our results showed that the 14-92 L/(m2×h) of the permeate flux was determined indicating a stabilized water productivity. In addition, the permeate water indicated a high removal rate (more than 99.99%) of turbidity and bacteria. Consequently, the system can provide a stabilized water production with safe permeate water quality during long-term operation. These findings exemplify an effective approach to decentralized drinking water treatment for developing countries.

  • https://doi.org/10.37675/jat.2021.7.1.72 인용