6533b82bfe1ef96bd128cd50
RESEARCH PRODUCT
Contact sites of peptide-oligoribonucleotide cross-links identified by a combination of peptide and nucleotide sequencing with MALDI MS.
Brigitte Wittmann-lieboldHenning UrlaubEva-christina MüllerBernd Thiedesubject
chemistry.chemical_classificationRibosomal ProteinsBinding SitesBase SequenceChemistryMolecular Sequence DataNucleic acid sequencePeptideRibosomal RNABiochemistryRibosomeAmino acidRNA BacterialBiochemistryBacterial ProteinsRibosomal proteinRNA RibosomalSpectrometry Mass Matrix-Assisted Laser Desorption-IonizationEscherichia coli30SAmino Acid SequencePeptide sequencedescription
We have investigated peptide–oligoribonucleotide complexes isolated from cross-linked Escherichia coli 30S ribosomal subunits in order to identify the contact sites of these complexes at the molecular level. For this purpose, reversed-phase (RP) HPLC-purified peptide–oligoribonucleotide complexes were submitted to N-terminal amino acid sequencing in order to determine the cross-linked peptide moiety and were analyzed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) for calculation of the nucleotide composition of the cross-linked complex. Subsequently, for nucleotide sequence information the complexes were partially hydrolyzed or treated with exonucleases and analyzed again by MALDI-MS. Applying this technique, we were able to identify the cross-linked oligoribonucleotide parts in contact with distinct peptide regions derived from ribosomal proteins S4, S7, S8, and S17 from E. coli.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1997-07-01 | Journal of protein chemistry |