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Systematic evaluation of the adaptability of the non-radioactive SUnSET assay to measure cardiac protein synthesis

Ravi, Venkatraman and Jain, Aditi and Ahamed, Faiz and Fathma, Nowrin and Desingu, Perumal Arumugam and Sundaresan, Nagalingam R (2018) Systematic evaluation of the adaptability of the non-radioactive SUnSET assay to measure cardiac protein synthesis. In: SCIENTIFIC REPORTS, 8 .

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Official URL: http://dx.doi.org/10.1038/s41598-018-22903-8

Abstract

Heart is a dynamic organ that undergoes remodeling in response to both physiological and pathological stimuli. One of the fundamental cellular processes that facilitates changes in the size and shape of this muscular organ is the protein synthesis. Traditionally changes in cardiac protein synthesis levels were measured by radiolabeled tracers. However, these methods are often cumbersome and suffer from radioactive risk. Recently a nonradioactive method for detecting protein synthesis under in vitro conditions called the Surface Sensing of Translation (SUnSET) was described in cell lines of mouse dendrites and T cells. In this work, we provide multiple lines of evidence that the SUnSET assay can be applied to reliably detect changes in protein synthesis both in isolated neonatal primary cardiomyocytes and heart. We successfully tracked the changes in protein synthesis by western blotting as well as immunohistochemical variants of the SUnSET assay. Applying the SUnSET assay, we measured the cardiac protein synthesis during the different ages of mice. Further, we successfully tracked the increase in cardiac protein synthesis during different stages of a well-established model for pathological hypertrophy. Overall, we propose SUnSET assay as a simple, reliable and robust method to measure protein synthesis in the cardiac milieu.

Item Type: Journal Article
Additional Information: Copy right for the article belong to NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Depositing User: Id for Latest eprints
Date Deposited: 02 Apr 2018 19:58
Last Modified: 03 Nov 2018 08:52
URI: http://eprints.iisc.ac.in/id/eprint/59437

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