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Functional Proteomics pp 197–224Cite as

Functional Proteomic Analysis to Characterize Signaling Crosstalk

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1871))

Abstract

The biological activities of a cell are determined by its response to external stimuli. The signals are transduced from either intracellular or extracellular milieu through networks of multi-protein complexes and post-translational modifications of proteins (PTMs). Most PTMs including phosphorylation, acetylation, ubiquitination, and SUMOylation, among others, modulate activities of proteins and regulate biological processes such as proliferation, differentiation, as well as host pathogen interaction. Conventionally, reverse genetics analysis and single molecule-based studies were employed to identify and characterize the function of PTMs and enzyme-substrate networks regulated by them. With the advent of high-throughput technologies, it is now possible to identify and quantify thousands of PTM sites in a single experiment. Here, we discuss recent advances in enrichment strategies of various PTMs. We also describe a method for the identification and relative quantitation of proteins using a tandem mass tag labeling approach combined with serial enrichment of phosphorylation, acetylation and succinylation using antibody enrichment strategy.

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Acknowledgements

The authors acknowledge Yenepoya (Deemed to be University) for access to mass spectrometry instrumentation facility. We also thank Karnataka Biotechnology and Information Technology Services (KBITS), Government of Karnataka, for the support to the Center for Systems Biology and Molecular Medicine at Yenepoya University under the Biotechnology Skill Enhancement Programme in Multiomics Technology (BiSEP GO ITD 02 MDA 2017). SMP is a recipient of INSPIRE Faculty Award from Department of Science and Technology (DST), Government of India.

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  1. Center for Systems Biology and Molecular Medicine, Yenepoya Research Centre, Yenepoya (Deemed to be University), Mangalore, India

    Sneha M. Pinto, Yashwanth Subbannayya & T. S. Keshava Prasad

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  1. Sneha M. Pinto
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  2. Yashwanth Subbannayya
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  3. T. S. Keshava Prasad
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Correspondence to T. S. Keshava Prasad .

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  1. Array Bridge Inc., St. Louis, MO, USA

    Xing Wang

  2. Biotech Support Group LLC, Monmouth Junction, NJ, USA

    Matthew Kuruc

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Pinto, S.M., Subbannayya, Y., Prasad, T.S.K. (2019). Functional Proteomic Analysis to Characterize Signaling Crosstalk. In: Wang, X., Kuruc, M. (eds) Functional Proteomics. Methods in Molecular Biology, vol 1871. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8814-3_14

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