Low temperature aqueous electrospray ionization mass spectrometry of noncovalent complexes

T. D. Veenstra, A. J. Tomlinson, L. Benson, Rajiv Kumar, S. Naylor

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

In the present study we describe conditions that permit the characterization of noncovalent protein-substrate complexes in aqueous solution by microspray electrospray ionization-mass spectrometry (ESI-MS), using a heated transfer capillary at low temperature (45 °C). Specifically, we examined the binding of calmodulin to two polypeptides; the calmodulin-binding domain of calmodulin-dependent protein kinase II (CamK-II) and melittin. Calmodulin, a well known calcium-binding protein, binds to a number of small amphipathic peptides in a calcium-dependent manner. Our results directly show that both peptides form equimolar complexes with calmodulin only in the presence of calcium. The stoichiometry necessary for the formation of each complex was 1:1:4 for calmodulin:peptide (melittin or CamK-II):Ca2+, respectively. Furthermore, it is demonstrated that the detection of the complex in ESI-MS is source temperature dependent.

Original languageEnglish (US)
Pages (from-to)580-584
Number of pages5
JournalJournal of the American Society for Mass Spectrometry
Volume9
Issue number6
DOIs
StatePublished - 1998

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Electrospray ionization
Electrospray Ionization Mass Spectrometry
Calmodulin
Mass spectrometry
Temperature
Melitten
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Peptides
Calcium
Calcium-Binding Proteins
Stoichiometry
Substrates
Proteins

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

Cite this

Low temperature aqueous electrospray ionization mass spectrometry of noncovalent complexes. / Veenstra, T. D.; Tomlinson, A. J.; Benson, L.; Kumar, Rajiv; Naylor, S.

In: Journal of the American Society for Mass Spectrometry, Vol. 9, No. 6, 1998, p. 580-584.

Research output: Contribution to journalArticle

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