Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay

Will A. Stanley, Kees Versluis, Carsten Schultz, Albert J.R. Heck, Matthias Wilmanns

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Sterol carrier protein 2 (SCP2) has been investigated by nearly native electrospray ionisation mass spectrometry in the presence of long chain fatty acyl CoAs (LCFA-CoAs) and carnitine derivatives of equivalent fatty acid chain length (LCFA-carnitines). Four SCP2 constructs were compared to examine the influence of the N-terminal presequence and the C-terminal peroxisomal targeting signal on ligand binding. Removal of N- or C-terminal residues did not influence ligand binding. The observation that LCFA-CoAs are high affinity ligands for SCP2 was confirmed, while LCFA-carnitines were demonstrated for the first time not to interact with SCP2. LCFA-CoAs formed non-covalent complexes with SCP2 of 2:1 and 1:1 stoichiometry, which could be dissociated by elevating the energy of the ions upon entrance to the mass spectrometer. A fluorescence-competition assay using Nile Red butyric acid confirmed the mass spectrometric observations in solution. The physiological significance of the lack of LCFA-carnitine binding by SCP2 is discussed.

Original languageEnglish (US)
Pages (from-to)50-58
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume461
Issue number1
DOIs
StatePublished - May 1 2007

Keywords

  • Electrospray mass spectrometry
  • Fatty acyl CoA
  • Fatty acyl carnitine
  • Nile Red butyric acid
  • Non-specific lipid transfer protein
  • Sterol carrier protein 2 (SCP2)

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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