A Comparative Proteomic Study of Human Skin Suction Blister Fluid From Healthy Individuals Using Immunodepletion and iTRAQ Labeling

J Proteome Res. 2012 Jul 6;11(7):3715-27. doi: 10.1021/pr3002035. Epub 2012 May 31.

Abstract

Aberrations in skin morphology and functionality can cause acute and chronic skin-related diseases that are the focus of dermatological research. Mechanically induced skin suction blister fluid may serve as a potential, alternative human body fluid for quantitative mass spectrometry (MS)-based proteomics in order to assist in the understanding of the mechanisms and causes underlying skin-related diseases. The combination of abundant-protein removal with iTRAQ technology and multidimensional fractionation techniques improved the number of identified protein groups. A relative comparison of a cohort of 8 healthy volunteers was thus recruited in order to assess the net variability encountered in a healthy scenario. The technology enabled the identification, to date, of the highest number of reported protein groups (739) with concomitant relative quantitative data for over 90% of all proteins with high reproducibility and accuracy. The use of iTRAQ 8-plex resulted in a 66% decrease in protein identifications but, despite this, provided valuable insight into interindividual differences of the healthy control samples. The geometric mean ratio was close to 1 with 95% of all ratios ranging between 0.45 and 2.05 and a calculated mean coefficient of variation of 15.8%, indicating a lower biological variance than that reported for plasma or urine. By applying a multistep sample processing, the obtained sensitivity and accuracy of quantitative MS analysis demonstrates the prospective value of the approach in future research into skin diseases.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blister / enzymology
  • Blister / metabolism*
  • Cohort Studies
  • Female
  • Health
  • Humans
  • Male
  • Middle Aged
  • Proteome / chemistry
  • Proteome / isolation & purification
  • Proteome / metabolism*
  • Reference Values
  • Ribonucleases / metabolism
  • Skin / enzymology
  • Skin / metabolism*
  • Staining and Labeling
  • Suction

Substances

  • Proteome
  • Ribonucleases