Lysine Methylation Mapping of Crenarchaeal DNA-Directed RNA Polymerases by Collision-Induced and Electron-Transfer Dissociation Mass Spectrometry
| Autor: | Felix Elortza, Nicola G. A. Abrescia, Mikel Azkargorta, Magdalena N. Wojtas |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Sulfolobus acidocaldarius
Archaeal Proteins Molecular Sequence Data ved/biology.organism_classification_rank.species Methylation Biochemistry Mass Spectrometry Sulfolobus Electron Transport chemistry.chemical_compound Species Specificity Transcription (biology) RNA polymerase Amino Acid Sequence Polymerase Sulfolobus shibatae Binding Sites Sequence Homology Amino Acid biology ved/biology Lysine Acetylation DNA-Directed RNA Polymerases General Chemistry biology.organism_classification Electron-transfer dissociation Protein Subunits chemistry biology.protein Electrophoresis Polyacrylamide Gel DNA Chromatography Liquid |
| Zdroj: | Journal of Proteome Research. 13:2637-2648 |
| ISSN: | 1535-3907 1535-3893 |
| DOI: | 10.1021/pr500084p |
| Popis: | Enzymatic machineries fundamental for information processing (e.g., transcription, replication, translation) in Archaea are simplified versions of their eukaryotic counterparts. This is clearly noticeable in the conservation of sequence and structure of corresponding enzymes (see for example the archaeal DNA-directed RNA polymerase (RNAP)). In Eukarya, post-translational modifications (PTMs) often serve as functional regulatory factors for various enzymes and complexes. Among the various PTMs, methylation and acetylation have been recently attracting most attention. Nevertheless, little is known about such PTMs in Archaea, and cross-methodological studies are scarce. We examined methylation and N-terminal acetylation of endogenously purified crenarchaeal RNA polymerase from Sulfolobus shibatae (Ssh) and Sulfolobus acidocaldarius (Sac). In-gel and in-solution protein digestion methods were combined with collision-induced dissociation (CID) and electron-transfer dissociation (ETD) mass spectrometry analysis. Overall, 20 and 26 methyl-lysines for S. shibatae and S. acidocaldarius were identified, respectively. Furthermore, two N-terminal acetylation sites for each of these organisms were assessed. As a result, we generated a high-confidence data set for the mapping of methylation and acetylation sites in both Sulfolobus species, allowing comparisons with the data previously obtained for RNAP from Sulfolobus solfataricus (Sso). We confirmed that all observed methyl-lysines are on the surface of the RNAP. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|