Details zur Publikation |
| Kategorie | Textpublikation |
| Referenztyp | Zeitschriften |
| DOI | 10.1016/j.talanta.2020.121711 |
| Volltext | Akzeptiertes Manuskript |
| Titel (primär) | Characterization of membrane-bound metalloproteins in the anaerobic ammonium-oxidizing bacterium “Candidatus Kuenenia stuttgartiensis” strain CSTR1 |
| Autor | Budhraja, R.; Karande, S.; Ding, C.; Ullrich, M.K.; Wagner, S.; Reemtsma, T.; Adrian, L. |
| Quelle | Talanta |
| Erscheinungsjahr | 2021 |
| Department | ISOBIO; UBT; ANA |
| Band/Volume | 223 |
| Heft | Part 2 |
| Seite von | art. 121711 |
| Sprache | englisch |
| Topic | T9 Healthy Planet T7 Bioeconomy |
| Keywords | Anammox; Metallome; Metalloproteomic; Blue native gel electrophoresis; Laser ablation; Desolvating nebulizer; ICP-MS |
| UFZ Querschnittsthemen | ProVIS; |
| Abstract | Membrane-bound metalloproteins are the basis of biological energy conservation via respiratory processes, however, their biochemical characterization is difficult. Here, we followed a gel-based proteomics and metallomics approach to identify membrane-associated metalloproteins in the anaerobic ammonium-oxidizing “Candidatus Kuenenia stuttgartiensis” strain CSTR1. Membrane-associated protein complexes were separated by two dimensional Blue Native/SDS gel electrophoresis and subunits were identified by mass spectrometry; protein-bound metal ions were quantified from the gel by connecting either a desolvating nebulizer system or laser ablation to inductively coupled plasma triple quadrupole mass spectrometry (ICP-QqQ-MS). We identified most protein complexes predicted to be involved in anaerobic ammonium oxidation and carbon fixation. The ICP-QqQ-MS data showed the presence of Fe and Zn in a wide range of high molecular weight protein complexes (230–800 kDa). Mo was prominently found in gel slices with proteins of a size of 500–650 kDa, whereas Ni was only found using the desolvating nebulizer system in the protein range of 350–500 kDa. The detected protein complexes and their metal content were consistent with genome annotations. Gel-based metalloproteomics is a sensitive and reliable approach for the characterization of metalloproteins and could be used to characterize many multimeric metalloprotein complexes in biological systems. |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=24075 |
| Budhraja, R., Karande, S., Ding, C., Ullrich, M.K., Wagner, S., Reemtsma, T., Adrian, L. (2021): Characterization of membrane-bound metalloproteins in the anaerobic ammonium-oxidizing bacterium “Candidatus Kuenenia stuttgartiensis” strain CSTR1 Talanta 223 (Part 2), art. 121711 10.1016/j.talanta.2020.121711 |
|