Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins

Takeshi Harayama, Hideo Shindou, Yoshihiro Kita, Eiji Otsubo, Kazushige Ikeda, Shoichi Chida, Timothy E. Weaver, Takao Shimizu

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The surfactant proteins (SPs), SP-B and SP-C, are important components of pulmonary surfactant involved in the reduction of alveolar surface tension. Quantification of SP-B and SP-C in surfactant drugs is informative for their quality control and the evaluation of their biological activity. Western blot analysis enabled the quantification of SP-B, but not SP-C, in surfactant drugs. Here, we report a new procedure involving chemical treatments and LC-MS to analyze SP-C peptides. The procedure enabled qualitative analysis of SP-C from different species with discrimination of the palmitoylation status and the artificial modifications that occur during handling and/or storage. In addition, the method can be used to estimate the total amount of SP-C in pulmonary surfactant drugs. The strategy described here might serve as a prototype to establish analytical methods for peptides that are extremely hydrophobic and behave like lipids. The new method provides an easy measurement of SP-C from various biological samples, which will help the characterization of various experimental animal models and the quality control of surfactant drugs, as well as diagnostics of human samples.

Original languageEnglish
Pages (from-to)1370-1379
Number of pages10
JournalJournal of Lipid Research
Volume56
Issue number7
DOIs
Publication statusPublished - 2015 Jul 1

Fingerprint

Surface-Active Agents
Protein C
Proteins
Pulmonary Surfactants
Quality Control
Pharmaceutical Preparations
Quality control
Lipoylation
Surface Tension
C-Peptide
Bioactivity
Surface tension
Animals
Animal Models
Western Blotting

Keywords

  • Drug therapy
  • Liquid chromatography
  • Lung
  • Mass spectrometry
  • Palmitoylation
  • Proteomics
  • Pulmonary surfactant
  • Surfactant drug

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

Harayama, T., Shindou, H., Kita, Y., Otsubo, E., Ikeda, K., Chida, S., ... Shimizu, T. (2015). Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins. Journal of Lipid Research, 56(7), 1370-1379. https://doi.org/10.1194/jlr.D060236

Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins. / Harayama, Takeshi; Shindou, Hideo; Kita, Yoshihiro; Otsubo, Eiji; Ikeda, Kazushige; Chida, Shoichi; Weaver, Timothy E.; Shimizu, Takao.

In: Journal of Lipid Research, Vol. 56, No. 7, 01.07.2015, p. 1370-1379.

Research output: Contribution to journalArticle

Harayama, T, Shindou, H, Kita, Y, Otsubo, E, Ikeda, K, Chida, S, Weaver, TE & Shimizu, T 2015, 'Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins', Journal of Lipid Research, vol. 56, no. 7, pp. 1370-1379. https://doi.org/10.1194/jlr.D060236
Harayama T, Shindou H, Kita Y, Otsubo E, Ikeda K, Chida S et al. Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins. Journal of Lipid Research. 2015 Jul 1;56(7):1370-1379. https://doi.org/10.1194/jlr.D060236
Harayama, Takeshi ; Shindou, Hideo ; Kita, Yoshihiro ; Otsubo, Eiji ; Ikeda, Kazushige ; Chida, Shoichi ; Weaver, Timothy E. ; Shimizu, Takao. / Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins. In: Journal of Lipid Research. 2015 ; Vol. 56, No. 7. pp. 1370-1379.
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