Abstract
Chlamydomonas is a model organism to study photosynthesis. Thylakoid membranes comprise several proteins belonging to photosystems I and II. In this chapter, we show the accurate proteomic measurements in thylakoid membranes. The chlorophyll-containing membrane protein complexes were precipitated using chloroform/methanol solution. These complexes were separated using two-dimensional gel electrophoresis, and the resolved spots were exercised from the gel matrix and digested with trypsin. These peptide fragments were separated by MALDI-TOF, and the isotopic masses were blasted to a MASCOT server to obtain the protein sequence. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF). The method discussed here would be a useful method for the separation and identification of thylakoid membrane proteins.
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Acknowledgments
RS is gratefully acknowledged for the financial support from DST NO.SR/SO/BB-34/2006, DST-FAST track NO SR/FT/L-89/2006 and DB-RGYI, Government of India. VY and SN thank CSIR and UGC India for a fellowship in the form of JRF and SRF. We thank Mark Hunter, Department of Chemistry and Biochemistry, Arizona State University, USA, for critical reading of this chapter. We greatly acknowledge the School of Life Sciences, UOH-CREBB, for providing the facility of MALDI-TOF mass spectrometry at the University of Hyderabad.
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Yadavalli, V., Nellaepalli, S., Subramanyam, R. (2011). Proteomic Analysis of Thylakoid Membranes. In: Carpentier, R. (eds) Photosynthesis Research Protocols. Methods in Molecular Biology, vol 684. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-925-3_14
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DOI: https://doi.org/10.1007/978-1-60761-925-3_14
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