Possibilities of subunit localization with fluorescent protein tags and electron microscopy examplified by a cyanobacterial NDH-1 study

Autor: Egbert J. Boekema, Mihaela Folea, Mariam Birungi, Eva-Mari Aro, Teruo Ogawa, Natalia Battchikova, Hualing Mi, Min Xu
Přispěvatelé: Groningen Biomolecular Sciences and Biotechnology, Electron Microscopy, Enzymology
Rok vydání: 2010
Předmět:
0106 biological sciences
Yellow fluorescent protein
THERMOSYNECHOCOCCUS-ELONGATUS
Protein subunit
Biophysics
Single particle analysis
CHLOROPLAST NAD(P)H DEHYDROGENASE
MITOCHONDRIAL COMPLEX-I
macromolecular substances
SYNECHOCYSTIS SP PCC-6803
Photosystem I
01 natural sciences
Biochemistry
Green fluorescent protein
law.invention
03 medical and health sciences
Subunit labeling
DOMAIN
Bacterial Proteins
law
NDH-1
Electron microscopy
LIVING CELLS
030304 developmental biology
0303 health sciences
ARCHITECTURE
Electron Transport Complex I
biology
IDENTIFICATION
Synechocystis
fungi
Cell Biology
Thermus thermophilus
Synechocystis 6803
biology.organism_classification
ARABIDOPSIS
Luminescent Proteins
Microscopy
Electron
Protein Subunits
THERMUS-THERMOPHILUS
biology.protein
Electron microscope
010606 plant biology & botany
Zdroj: Biochimica et Biophysica Acta-Bioenergetics, 1797(8), 1681-1686
ISSN: 0006-3002
0005-2728
Popis: Cyanobacterial NDH-1 is a multisubunit complex involved in proton translocation, cyclic electron flow around photosystem I and CO(2) uptake. The function and location of several of its small subunits are unknown. In this work, the location of the small subunits NdhL, -M, -N, -O and CupS of Synechocystis 6803 NDH-1 was established by electron microscopy (EM) and single particle analysis. To perform this, the subunits were enlarged by fusion with the YFP protein. After classification of projections, the position of the YFP tag was revealed; all five subunits are integrated in the membrane domain. The results on NDH-1 demonstrate that a GFP tag can be revealed after data processing of EM data sets of moderate size, thus showing that this way of labeling is a fast and reliable way for subunit mapping in multisubunit complexes after partial purification. (C) 2010 Elsevier B.V. All rights reserved.
Databáze: OpenAIRE
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