Non-invasive proteomic analysis of human skin keratins: Screening of methionine oxidation in keratins by mass spectrometry

研究成果: Article査読

16 被引用数 (Scopus)

抄録

Keratins are the main constituent of human skin and have been identified as major oxidative target proteins. However, there has been a lack of studies aimed at identifying the oxidation sites of keratins because of the difficulties associated with their insolubility and handling. Here, we introduce a mass spectrometry (MS)-based proteomic methodology to screen oxidative modifications in human skin keratins. Human skin proteins were obtained non-invasively by tape stripping and solubilized in SDS buffer, followed by purification and digestion using the modified filter-aided sample preparation method. The tryptic peptides were then analyzed by MALDI-TOF/MS, LC-ESI/MS, and MS/MS. PMF analyses have identified keratins K1 and K10 as the major proteins of human skin. Met 259, Met 262, Met 296, and Met 469, located in the α-helical rod domain of K1, were the most susceptible sites to oxidation induced by hydrogen peroxide in vitro and in vivo. Our results indicate a potential use of the identified methionine residues as biomarkers of oxidative skin damage. The present methodology is the first MS-based approach to detecting oxidative modifications in keratins obtained directly from human skin and can be easily applied to the monitoring of other keratin modifications in various skin conditions.

本文言語English
ページ(範囲)435-449
ページ数15
ジャーナルJournal of Proteomics
75
2
DOI
出版ステータスPublished - 2011 12 21

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

フィンガープリント 「Non-invasive proteomic analysis of human skin keratins: Screening of methionine oxidation in keratins by mass spectrometry」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル