MALDI-MS detection of noncovalent interactions of single stranded DNA with Escherichia coli single-stranded DNA-binding protein

Metadata Label Value
Author(s) Chen, Fan, Mädler, Stefanie, Weidmann, Simon, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
Full Text Search SFX for a Full-Text version of this document
Import to Mendeley

Detailed Information

Metadata Field Content
Title MALDI-MS detection of noncovalent interactions of single stranded DNA with Escherichia coli single-stranded DNA-binding protein
Author(s) Chen, Fan
Mädler, Stefanie
Weidmann, Simon
Zenobi, Renato
Journal or Series Title Journal of mass spectrometry
Volume Number 47
Issue Number 5
Start Page 560
End Page 566
ISSN 1076-5174
Publisher Wiley
Publication Place Chichester
Publication Date 2012-05
Keyword(s) Single-stranded DNA binding protein
Protein-DNA interactions
Chemical cross-linking
Noncovalent complexes
Abstract The Escherichia coli single-stranded DNA binding protein (SSB) selectively binds single-stranded (ss) DNA and participates in the process of DNA replication, recombination and repair. Different binding modes have previously been observed in SSBssDNA complexes, due to the four potential binding sites of SSB. Here, chemical cross-linking, combined with high-mass matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS), is used to determine the stoichiometry of the SSBssDNA complex. SSB forms a stable homotetramer in solution, but only the monomeric species (m/z 19?100) can be detected with standard MALDI-MS. With chemical cross-linking, the quaternary structure of SSB is conserved, and the tetramer (m/z 79?500) was observed. We found that ssDNA also functions as a stabilizer to conserve the quaternary structure of SSB, as evidenced by the detection of a SSBssDNA complex at m/z 94?200 even in the absence of chemical cross-linking. The stability of the SSBssDNA complex with MALDI strongly depends on the length and strand of oligonucleotides and the stoichiometry of the SSBssDNA complex, which could be attributed to electrostatic interactions that are enhanced in the gas phase. The key factor affecting the stoichiometry of the SSBssDNA complex is how ssDNA binds to SSB, rather than the protein-to-DNA ratio. This further suggests that detection of the complex by MALDI is a result of specific binding, and not due to non-specific aggregation in the MALDI plume.
DOI 10.1002/jms.2989
Additional Notes Received 25 December 2011, Revised 5 March 2012, Accepted 14 March 2012
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
Organisational Unit(s)
NEBIS System Number 001410098
Source Database ID WOS-000303439700004
Description File Name MIME Type Size
No details could be found
There are no links available for this record.
This record has not been viewed during this period

  author = "Chen, Fan and M{\"{a}}dler, Stefanie and Weidmann, Simon and Zenobi, Renato",
  title = "{M}{A}{L}{D}{I}-{M}{S} detection of noncovalent interactions of single stranded {D}{N}{A} with {E}scherichia coli single-stranded {D}{N}{A}-binding protein",
  journal = "Journal of mass spectrometry",
  year = 2012,
  volume = "47",
  number = "5",
  pages = "560--566",
  month = may,

E-Citations record created: Tue, 29 May 2012, 07:34:59 CET