Isolation and purification assay of ex vivo photosystem II D1 protein toward integrated biointeraction analysis

• Published in: Analytical and bioanalytical chemistry Vol. 390; no. 4; pp. 1195 - 1202
• Main Authors: Muktiono, B, Schulten, C, Heemken, O, Gandrass, J, Prange, A, Schnabl, H, Cerboncini, C
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-02-2008; Springer-Verlag
• Subjects: Analytical Chemistry; Biochemistry; Blotting, Western; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatography, Gel; D1 protein; Electrophoresis, Polyacrylamide Gel; Food Science; Laboratory Medicine; Monitoring/Environmental Analysis; Oxygen-evolving complex; Photosystem II Protein Complex - chemistry; Photosystem II Protein Complex - isolation & purification; Photosystem II Protein Complex - metabolism; Protein Binding; Spectrometry, Mass, Electrospray Ionization; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Technical Note; thylakoids; Oxygen-evolving complex; D1 protein; Thylakoids
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-007-1781-2

Protein extracts of photosystem II were prepared from leaf chloroplasts of different plant species by fast and nondenaturing methods. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot analysis of the proteins obtained showed that the extracts were enriched by D1 proteins, which appeared putatively in association with the 33-kDa oxygen-evolving-complex subunits. In further isolation steps D1 proteins were purified using salt-gradient chromatography (fast protein liquid chromatography) and characterized by western blot and mass spectrometry.