Driving Forces of Proteasome-catalyzed Peptide Splicing in Yeast and Humans
| Autor: | Alexander Kloss, Burkhardt Dahlmann, Petra Henklein, Katharina Janek, Andrean Goede, Christin Keller, Michele Mishto, Agathe Niewienda, Sabrina Gohlke, Kathrin Textoris Taube, Cordula Enenkel, Peter M. Kloetzel |
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| Rok vydání: | 2012 |
| Předmět: |
Proteasome Endopeptidase Complex
Molecular Sequence Data education Saccharomyces cerevisiae Peptide Biochemistry Analytical Chemistry Protein splicing Humans Protein Splicing Amino Acid Sequence Binding site Molecular Biology Peptide sequence Cell Line Transformed chemistry.chemical_classification B-Lymphocytes biology Research Active site biology.organism_classification Proteasome chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization RNA splicing Biocatalysis biology.protein Peptides Chromatography Liquid |
| Zdroj: | Molecular & Cellular Proteomics. 11:1008-1023 |
| ISSN: | 1535-9476 |
| Popis: | Proteasome-catalyzed peptide splicing (PCPS) represents an additional activity of mammalian 20S proteasomes recently identified in connection with antigen presentation. We show here that PCPS is not restricted to mammalians but that it is also a feature of yeast 20S proteasomes catalyzed by all three active site β subunits. No major differences in splicing efficiency exist between human 20S standard- and immuno-proteasome or yeast 20S proteasome. Using H(2)(18)O to monitor the splicing reaction we also demonstrate that PCPS occurs via direct transpeptidation that slightly favors the generation of peptides spliced in cis over peptides spliced in trans. Splicing efficiency itself is shown to be controlled by proteasomal cleavage site preference as well as by the sequence characteristics of the spliced peptides. By use of kinetic data and quantitative analyses of PCPS obtained by mass spectrometry we developed a structural model with two PCPS binding sites in the neighborhood of the active Thr1. |
| Databáze: | OpenAIRE |
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