Abstract
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.
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Urlaub, H., Thiede, B., Müller, EC. et al. Contact Sites of Peptide–Oligoribonucleotide Cross-Links Identified by a Combination of Peptide and Nucleotide Sequencing with MALDI MS. J Protein Chem 16, 375–383 (1997). https://doi.org/10.1023/A:1026380504377
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DOI: https://doi.org/10.1023/A:1026380504377