Using ESI-MS to probe protein structure by site-specific noncovalent attachment of 18-crown-6

Tony Ly, Ryan R. Julian*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

A new method for probing the equilibrium structures and folding states of proteins utilizing electrospray ionization mass spectrometry is described. Protein structure is explored as a function of side-chain availability as determined by a specific interaction between lysine and 18-crown-6 ether (18C6). Various intramolecular interactions are competitive with the lysine/18C6 interaction and can prevent noncovalent attachment of 18C6. Changes to protein structure modify these inhibiting intramolecular interactions, which leads to a change in the number of 18C6s that attach to the protein. Experiments conducted with cytochrome c, ubiquitin, and melittin reveal that the method is sensitive to changes in both tertiary and secondary structure. In addition, the structure of each charge state can be examined independently. Experiments can be performed under conditions where the pH and amount of organic cosolvent are varied. Control experiments conducted with pentalysine, which lacks structural organization, are also presented.

Original languageEnglish
Pages (from-to)1209-1215
Number of pages7
JournalJournal of the American Society for Mass Spectrometry
Volume17
Issue number9
DOIs
Publication statusPublished - Sep 2006

Keywords

  • IONIZATION MASS-SPECTROMETRY
  • CHARGE-STATE DISTRIBUTIONS
  • ALPHA-HELIX FORMATION
  • HYDROGEN-EXCHANGE
  • CYTOCHROME-C
  • LIGAND INTERACTIONS
  • CIRCULAR-DICHROISM
  • UBIQUITIN
  • MELITTIN
  • METHANOL

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