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




TekijätBirungi M, Folea M, Battchikova N, Xu M, Mi HL, Ogawa T, Aro EM, Boekema EJ

KustantajaELSEVIER SCIENCE BV

Julkaisuvuosi2010

JournalBBA - Bioenergetics

Tietokannassa oleva lehden nimiBIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS

Lehden akronyymiBBA-BIOENERGETICS

Numero sarjassa8

Vuosikerta1797

Numero8

Aloitussivu1681

Lopetussivu1686

Sivujen määrä6

ISSN0005-2728

DOIhttps://doi.org/10.1016/j.bbabio.2010.06.004


Tiivistelmä
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.



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