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 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Mass spectrometry
Assays
Mass Spectrometry
Carnitine
Ligands
Electrospray ionization
Butyric Acid
Electrospray Ionization Mass Spectrometry
Mass spectrometers
Chain length
Stoichiometry
sterol carrier proteins
Fatty Acids
Fluorescence
Ions
Derivatives

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay. / Stanley, Will A.; Versluis, Kees; Schultz, Carsten; Heck, Albert J R; Wilmanns, Matthias.

In: Archives of Biochemistry and Biophysics, Vol. 461, No. 1, 01.05.2007, p. 50-58.

Research output: Contribution to journalArticle

Stanley, Will A. ; Versluis, Kees ; Schultz, Carsten ; Heck, Albert J R ; Wilmanns, Matthias. / Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay. In: Archives of Biochemistry and Biophysics. 2007 ; Vol. 461, No. 1. pp. 50-58.
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