Analysis of the chloroplast protein complexes by blue-native polyacrylamide gel electrophoresis (BN-PAGE)

verfasst von: Marion Kügler, Lothar Jänsch, Volker Kruft, Udo Schmitz, Hans-Peter Braun
Abstract: Blue-native polyacrylamide gel electrophoresis (BN-PAGE) is a powerful procedure for the separation and characterization of the protein complexes from mitochondria. Membrane proteins are solubilized in the presence of aminocaproic acid and n-dodecylmaltoside and Coomassie-dyes are utilized before electrophoresis to introduce a charge shift on proteins. Here, we report a modification of the procedure for the analysis of chloroplast protein complexes. The two photosystems, the light-harvesting complexes, the ATP synthase, the cytochrome b₆f complex and the ribulose-bisphosphate carboxylase/oxygenase are well resolved. Analysis of the protein complexes on a second gel dimension under denaturing conditions allows separation of more than 50 different proteins which are part of chloroplast multi-subunit enzymes. The resolution capacity of the blue-native gels is very high if compared to 'native green gel systems' published previously. N-terminal amino acid sequences of single subunits can be directly determined by cyclic Edman degradation as demonstrated for eight proteins. Analysis of chloroplast protein complexes by blue-native gel electrophoresis will allow the generation of 'protein maps' from different species, tissues and developmental stages or from mutant organelles. Further applications of blue-native gel electrophoresis are discussed.
Organisationseinheit(en): Abteilung Pflanzenmolekularbiologie und Pflanzenproteomik
Externe Organisation(en): Applied Biosystems GmbH
Typ: Artikel
Journal: Photosynthesis Research
Band: 53
Seiten: 35-44
Anzahl der Seiten: 10
ISSN: 0166-8595
Publikationsdatum: 07.1997
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Biochemie, Pflanzenkunde, Zellbiologie
Elektronische Version(en): https://doi.org/10.1023/A:1005882406718 (Zugang: Unbekannt)