title: A comparative proteomic study of human skin suction blister fluid from healthy individuals using immunodepletion and iTRAQ labeling.
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.
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: André C
authors.initials: AC
authors.lastname: Müller
authors.forename: Florian P
authors.initials: FP
authors.lastname: Breitwieser
authors.forename: Heinz
authors.initials: H
authors.lastname: Fischer
authors.forename: Christopher
authors.initials: C
authors.lastname: Schuster
authors.forename: Oliver
authors.initials: O
authors.lastname: Brandt
authors.forename: Jacques
authors.initials: J
authors.lastname: Colinge
authors.forename: Giulio
authors.initials: G
authors.lastname: Superti-Furga
authors.forename: Georg
authors.initials: G
authors.lastname: Stingl
authors.forename: Adelheid
authors.initials: A
authors.lastname: Elbe-Bürger
authors.forename: Keiryn L
authors.initials: KL
authors.lastname: Bennett
mesh_terms: D000328:Adult; D001768:Blister / Q000201:enzymology / Q000378:metabolism*; D015331:Cohort Studies; D005260:Female; D006262:Health; D006801:Humans; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012016:Reference Values; D012260:Ribonucleases / Q000378:metabolism; D012867:Skin / Q000201:enzymology / Q000378:metabolism*; D013194:Staining and Labeling; D013396:Suction
chemical_list: D020543:Proteome; D012260:Ribonucleases