title: Possibilities of subunit localization with fluorescent protein tags and electron microscopy examplified by a cyanobacterial NDH-1 study.
abstract: Cyanobacterial NDH-1 is a multisubunit complex involved in proton translocation, cyclic electron flow around photosystem I and CO2 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.
authors.affiliation: Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Mariam
authors.initials: M
authors.lastname: Birungi
authors.forename: Mihaela
authors.initials: M
authors.lastname: Folea
authors.forename: Natalia
authors.initials: N
authors.lastname: Battchikova
authors.forename: Min
authors.initials: M
authors.lastname: Xu
authors.forename: Hualing
authors.initials: H
authors.lastname: Mi
authors.forename: Teruo
authors.initials: T
authors.lastname: Ogawa
authors.forename: Eva-Mari
authors.initials: EM
authors.lastname: Aro
authors.forename: Egbert J
authors.initials: EJ
authors.lastname: Boekema
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D042967:Electron Transport Complex I / Q000737:chemistry*; D008164:Luminescent Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D021122:Protein Subunits; D046939:Synechocystis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D021122:Protein Subunits; C054562:yellow fluorescent protein, Bacteria; D042967:Electron Transport Complex I