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Subcellular localization of an ATPase in anammox bacteria using proteomics and immunogold electron microscopy

Artikel i vetenskaplig tidskrift
Författare R. Karlsson
A. Karlsson
Ola Bäckman
Bengt R Johansson
Stefan Hulth
Publicerad i Fems Microbiology Letters
Volym 354
Nummer/häfte 1
Sidor 10-18
ISSN 0378-1097
Publiceringsår 2014
Publicerad vid Institutionen för biomedicin
Institutionen för kemi och molekylärbiologi
Sidor 10-18
Språk en
Länkar dx.doi.org/10.1111/1574-6968.12425
Ämnesord ATPase, anammoxosome, linear epitope, immunogold EM, anammox bacteria, ANAEROBIC AMMONIUM OXIDATION, CANDIDATUS KUENENIA STUTTGARTIENSIS, OXIDIZING BACTERIA, INTRACYTOPLASMIC COMPARTMENT, CELL, PROTEIN, MICROORGANISMS, PLAN, Microbiology, MICROBIOLOGY
Ämneskategorier Mikrobiologi inom det medicinska området

Sammanfattning

Anaerobic ammonium oxidation (anammox) has received significant attention during optimization of waste-water treatment and constitutes an important pathway for the removal of bioavailable nitrogen from natural environments. Studies of key catabolic enzymes indicate that the anammox reaction takes place inside the anammoxosome, an organelle-like membranous compartment of anammox bacteria. The anammoxosome has also been suggested as a site for ATP synthesis. A lipid-based protein immobilization technique, previously used to identify proteins essential for the anammox reaction, was in this study used to select linear epitopes for antibodies specifically targeted against an identified ATPase. The approach of using proteomics and bioinformatics as tools for selecting antibody targets for immunolocalization provides an important alternative to traditional methods for selection of specific antibodies. Immunogold electron microscopy and statistical evaluations indicated that the antibodies against the ATPase were exclusively found associated with the anammoxosome membrane. This provides strong evidence for ATP synthesis by an intracellular proton motive force in anammox bacteria. Within prokaryotes, an ATP synthase associated with an intracellular compartment is a feature unique for anammox bacteria.

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