PMID:25962914

Citation	de Almeida, NM, Neumann, S, Mesman, RJ, Ferousi, C, Keltjens, JT, Jetten, MS, Kartal, B and van Niftrik, L (2015) Immunogold Localization of Key Metabolic Enzymes in the Anammoxosome and on the Tubule-Like Structures of Kuenenia stuttgartiensis. J. Bacteriol. 197:2432-41
Abstract	Anaerobic ammonium-oxidizing (anammox) bacteria oxidize ammonium with nitrite as the terminal electron acceptor to form dinitrogen gas in the absence of oxygen. Anammox bacteria have a compartmentalized cell plan with a central membrane-bound "prokaryotic organelle" called the anammoxosome. The anammoxosome occupies most of the cell volume, has a curved membrane, and contains conspicuous tubule-like structures of unknown identity and function. It was suggested previously that the catalytic reactions of the anammox pathway occur in the anammoxosome, and that proton motive force was established across its membrane. Here, we used antibodies raised against five key enzymes of the anammox catabolism to determine their cellular location. The antibodies were raised against purified native hydroxylamine oxidoreductase-like protein kustc0458 with its redox partner kustc0457, hydrazine dehydrogenase (HDH; kustc0694), hydroxylamine oxidase (HOX; kustc1061), nitrite oxidoreductase (NXR; kustd1700/03/04), and hydrazine synthase (HZS; kuste2859-61) of the anammox bacterium Kuenenia stuttgartiensis. We determined that all five protein complexes were exclusively located inside the anammoxosome matrix. Four of the protein complexes did not appear to form higher-order protein organizations. However, the present data indicated for the first time that NXR is part of the tubule-like structures, which may stretch the whole length of the anammoxosome. These findings support the anammoxosome as the locus of catabolic reactions of the anammox pathway.
Links	PubMed PMC4524196 Online version:10.1128/JB.00186-15
Keywords	Ammonium Compounds/metabolism; Anaerobiosis; Bacteria/cytology; Bacteria/enzymology; Bacteria/metabolism; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Gene Expression Regulation, Bacterial/physiology; Immunohistochemistry; Organelles/physiology; Oxidation-Reduction
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9BACT:Q1PW30		GO:0044222: anammoxosome	ECO:0000314:		C		Fig. 6 stuttgartiensis cells incubated with antibodies against HDH. Thin sections were incubated with 1:100-diluted anti-HDH (A and B), 1:50-diluted preimmune serum (C), or 1:100-diluted anti-HDH (D). Gold particles were visible only inside the anammoxosome"	complete CACAO 11163
9BACT:Q1PX48		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 5- “ Thin sections were incubated with 1:300-diluted anti-HOX (A and B) or 1:100-diluted anti-HOX (C and D). Gold particles were visible only inside the anammoxosome. HOX appeared to be preferentially localized in the proximity of the anammoxosome membrane.”	complete CACAO 11162
9BACT:Q1Q0T3		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 7 -“K. stuttgartiensis cells incubated with affinity-purified antibodies against HZS. Thin sections were incubated with 1:200-diluted anti-HZS (A to C) or 1:300-diluted anti-HZS (D). Gold particles were visible only inside the anammoxosome. “	complete CACAO 11153
9BACT:Q1Q0T2		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 7- “K. stuttgartiensis cells incubated with antibodies against HZS. Thin sections were incubated with 1:100-diluted anti-HZS (A and B), 1:50-diluted preimmune serum (C), or 1:100-diluted anti-HZS (D). Gold particles were visible only inside the anammoxosome.”	complete CACAO 11154
9BACT:Q1Q0T4		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 7 - “K. stuttgartiensis cells incubated with affinity-purified antibodies against HZS. Thin sections were incubated with 1:200-diluted anti-HZS (A to C) or 1:300-diluted anti-HZS (D). Gold particles were visible only inside the anammoxosome. “	complete CACAO 11155
9BACT:Q1PVE0		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 4 - “K. stuttgartiensis cells incubated with antibodies against kustc0457/58. Thin sections were incubated with 1:150-diluted anti-kustc0457/58 (A and B), 1:50-diluted preimmune serum (C), or 1:100-diluted anti-kustc0457/58 (D). Gold particles were visible only inside the anammoxosome, preferentially located toward the inner side of the anammoxosome membrane. Incubation with preimmune serum led to the detection of a few gold particles throughout the cell.”	complete CACAO 11160
9BACT:Q1PVE1		GO:0044222: anammoxosome	ECO:0000314:		C		Fig 4 - "K. stuttgartiensis cells incubated with antibodies against kustc0457/58. Thin sections were incubated with 1:150-diluted anti-kustc0457/58 (A and B), 1:50-diluted preimmune serum (C), or 1:100-diluted anti-kustc0457/58 (D). Gold particles were visible only inside the anammoxosome, preferentially located toward the inner side of the anammoxosome membrane. Incubation with preimmune serum led to the detection of a few gold particles throughout the cell.”	complete CACAO 11161
9BACT:Q1PZD8		GO:0044222: anammoxosome	ECO:0000314:		C		Results: “In contrast, anti-NXR was very specifically observed at the tubule-like structures in the anammoxosome (Fig. 8) in both transverse (Fig. 8C) and longitudinal (Fig. 8A, B, and D) sectioned tubules.”	complete CACAO 11156
9BACT:Q1PZD5		GO:0044222: anammoxosome	ECO:0000314:		C		Results: “In contrast, anti-NXR was very specifically observed at the tubule-like structures in the anammoxosome (Fig. 8) in both transverse (Fig. 8C) and longitudinal (Fig. 8A, B, and D) sectioned tubules.”	complete CACAO 11157
9BACT:Q1PZD4		GO:0044222: anammoxosome	ECO:0000314:		C		Results: “anti-NXR was very specifically observed at the tubule-like structures in the anammoxosome (Fig. 8) in both transverse (Fig. 8C) and longitudinal (Fig. 8A, B, and D) sectioned tubules.”	complete CACAO 11158

References

See Help:References for how to manage references in GONUTS.