Proteomic analysis of thylakoid membranes.
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.
Yadavalli
Venkateswarlu
V
Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, India.
Nellaepalli
Sreedhar
S
Subramanyam
Rajagopal
R
D053000:Analytic Sample Preparation Methods; D005591:Chemical Fractionation; D016825:Chlamydomonas reinhardtii / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D002734:Chlorophyll / Q000378:metabolism; D057225:Data Mining; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D007525:Isoelectric Focusing; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020524:Thylakoids / Q000737:chemistry* / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D010940:Plant Proteins; D002734:Chlorophyll; D014357:Trypsin