Open AccessSubcellular localization of an ATPase in anammox bacteria using proteomics and immunogold electron microscopy
Author(s) -
Karlsson Roger,
Karlsson Anders,
Bäckman Ola,
Johansson Bengt R.,
Hulth Stefan
Publication year - 2014
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12425
Subject(s) - anammox , immunogold labelling , bacteria , biochemistry , atpase , biology , atp synthase , intracellular , chemiosmosis , organelle , proteomics , enzyme , microbiology and biotechnology , chemistry , antibody , genetics , denitrifying bacteria , organic chemistry , gene , nitrogen , immunology , denitrification
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.