Cross-linked ammonia monooxygenase complex in BN-PAGE gel from Nitrososphaera viennensis
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ABSTRACT: A band containing the ammonia monooxygenase complex from Nitrosospheara viennensis was cut from a Blue Native PAGE gel and cross linked using DSSO.
Project description:Ammonia-oxidizing archaea (AOA) are among the most abundant microorganisms and key players in the global nitrogen and carbon cycles. They share a common energy metabolism but represent a heterogeneous group with respect to their environmental distri- bution and adaptions, growth requirements, and genome contents. We report here the genome and proteome of Nitrososphaera viennensis EN76, the type species of the archaeal class Nitrososphaeria of the phylum Thaumarchaeota encompassing all known AOA. N. viennensis is a soil organism with a 2.52-Mb genome and 3,123 predicted protein-coding genes. Proteomic analysis revealed that nearly 50% of the predicted genes were translated under standard laboratory growth conditions. Comparison with genomes of closely related species of the predominantly terrestrial Nitrososphaerales as well as the more streamlined marine Nitrosopumilales (Candidatus order) and the acidophile Nitrosotalea devanaterra revealed a core genome of AOA comprising 860 genes, which allowed for the reconstruction of central metabolic pathways common to all known AOA and expressed in the N. viennensis and Nitrosopelagicus brevis proteomes. Concomitantly, we were able to identify candidate proteins for as yet unidentified crucial steps in central metabolisms. In addition to unraveling aspects of core AOA metabolism, we identified specific metabolic innovations associated with the Nitrososphaerales mediating growth and survival in the soil milieu, including the capacity for biofilm formation, cell surface modifications and cell adhesion, and carbohydrate conversions as well as detoxification of aromatic compounds and drugs.
Project description:Pure cultures of N. viennensis were subjected to copper limitation using the copper chelator TETA. Two samples that were subjected to copper limiation for approximately two weeks were used for protein extraction and subsequent proteomic analysis.
Project description:Information on the files related to the respective experiments can be found in the metadata file titled “Metadata_for_Nviennensis_Carbon_Experiment.xlsx”. The ammonia-oxidizing archaeon Nitrososphaera viennensis was subjected to various levels of carbon limitation based on predictions from a thermodynamic model. Catalase or pyruvate was used as a reactive oxygen species scavenger. A combined extraction of proteins and metabolites was performed in each condition to evaluate the cellular response to carbon limitation. The proteomic data associated with the project can be found here.
Project description:human OGDH (E1o) and DHTKD1 (E1a) were cross-linked with DLST (E2o). The samples were analyzed by LC-MS/MS in order to elucidate the interaction sites between these two E1-E2 subcomplexes.
Project description:CL-MS OF hE1a (DHTKD1) or G729R hE1a with hE2o to identify the possible difference in the interactions between hE1a-hE2o and G729R hE1a-hE2o
Project description:In this project, we compare the abundancies of cross-links in wild type and mutant iNOSoxyFMN-CaM complexes at specific interdomain interacting site and map the identified sites onto top AlphaFold 2 models for better understanding of iNOS protein dynamics.