A1 Refereed original research article in a scientific journal
Possibilities of subunit localization with fluorescent protein tags and electron microscopy examplified by a cyanobacterial NDH-1 study
Authors: Birungi M, Folea M, Battchikova N, Xu M, Mi HL, Ogawa T, Aro EM, Boekema EJ
Publisher: ELSEVIER SCIENCE BV
Publication year: 2010
Journal: BBA - Bioenergetics
Journal name in source: BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS
Journal acronym: BBA-BIOENERGETICS
Number in series: 8
Volume: 1797
Issue: 8
First page : 1681
Last page: 1686
Number of pages: 6
ISSN: 0005-2728
DOI: https://doi.org/10.1016/j.bbabio.2010.06.004
Abstract
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.
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.
UTU Research Portal