Show simple item record

dc.contributor.authorPérez-Pérez, Rafael
dc.contributor.authorLopez, Juan Antonio 
dc.contributor.authorGarcía-Santos, Eva
dc.contributor.authorCamafeita, Emilio 
dc.contributor.authorGomez-Serrano, Maria 
dc.contributor.authorOrtega-Delgado, Francisco J
dc.contributor.authorRicart, Wifredo
dc.contributor.authorFernández-Real, José M
dc.contributor.authorPeral, Belén
dc.identifier.citationPLoS One. 2012; 7(1):e30326es_ES
dc.description.abstractBACKGROUND: Protein expression studies based on the two major intra-abdominal human fat depots, the subcutaneous and the omental fat, can shed light into the mechanisms involved in obesity and its co-morbidities. Here we address, for the first time, the identification and validation of reference proteins for data standardization, which are essential for accurate comparison of protein levels in expression studies based on fat from obese and non-obese individuals. METHODOLOGY AND FINDINGS: To uncover adipose tissue proteins equally expressed either in omental and subcutaneous fat depots (study 1) or in omental fat from non-obese and obese individuals (study 2), we have reanalyzed our previously published data based on two-dimensional fluorescence difference gel electrophoresis. Twenty-four proteins (12 in study 1 and 12 in study 2) with similar expression levels in all conditions tested were selected and identified by mass spectrometry. Immunoblotting analysis was used to confirm in adipose tissue the expression pattern of the potential reference proteins and three proteins were validated: PARK7, ENOA and FAA. Western Blot analysis was also used to test customary loading control proteins. ENOA, PARK7 and the customary loading control protein Beta-actin showed steady expression profiles in fat from non-obese and obese individuals, whilst FAA maintained steady expression levels across paired omental and subcutaneous fat samples. CONCLUSIONS: ENOA, PARK7 and Beta-actin are proper reference standards in obesity studies based on omental fat, whilst FAA is the best loading control for the comparative analysis of omental and subcutaneous adipose tissues either in obese and non-obese subjects. Neither customary loading control proteins GAPDH and TBB5 nor CALX are adequate standards in differential expression studies on adipose tissue. The use of the proposed reference proteins will facilitate the adequate analysis of proteins differentially expressed in the context of obesity, an aim difficult to achieve before this study.es_ES
dc.description.sponsorshipThis work was supported by Grants SAF-2009-10461 and SAF-2008-02073 from the MICINN, and from the Fundacion Mutua Madrileña, Spain (BP). The CNIC is supported by MICINN and Pro CNIC Foundation. CIBERobn is an initiative from the Instituto de Salud Carlos III.es_ES
dc.publisherPublic Library of Sciencees_ES
dc.relation.isversionofPublisher's versiones_ES
dc.subject.meshActins es_ES
dc.subject.meshAdult es_ES
dc.subject.meshBiomarkers es_ES
dc.subject.meshBiomarkers, Tumor es_ES
dc.subject.meshBlotting, Westernes_ES
dc.subject.meshDNA-Binding Proteins es_ES
dc.subject.meshElectrophoresis, Gel, Two-Dimensional es_ES
dc.subject.meshFemale es_ES
dc.subject.meshHumans es_ES
dc.subject.meshImmunohistochemistry es_ES
dc.subject.meshIntracellular Signaling Peptides and Proteins es_ES
dc.subject.meshMiddle Aged es_ES
dc.subject.meshObesity es_ES
dc.subject.meshOmentum es_ES
dc.subject.meshOncogene Proteins es_ES
dc.subject.meshPhosphopyruvate Hydratase es_ES
dc.subject.meshProtein Deglycase DJ-1 es_ES
dc.subject.meshProteins es_ES
dc.subject.meshProteome es_ES
dc.subject.meshProteomics es_ES
dc.subject.meshReference Standards es_ES
dc.subject.meshReproducibility of Results es_ES
dc.subject.meshSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionizationes_ES
dc.subject.meshSubcutaneous Fat es_ES
dc.subject.meshTumor Suppressor Proteins es_ES
dc.titleUncovering suitable reference proteins for expression studies in human adipose tissue with relevance to obesityes_ES
dc.rights.licenseAtribución 4.0 Internacional*
dc.contributor.funderMinisterio de Ciencia e Innovación (España)es_ES
dc.contributor.funderFundación Mutua Madrileñaes_ES
dc.contributor.funderFundación ProCNICes_ES
dc.contributor.funderInstituto de Salud Carlos III - ISCIIIes_ES
dc.identifier.journalPloS onees_ES
dc.repisalud.orgCNICCNIC::Unidades técnicas::Proteómica / Metabolómicaes_ES

Files in this item

Acceso Abierto

This item appears in the following Collection(s)

Show simple item record

Atribución 4.0 Internacional
This item is licensed under a: Atribución 4.0 Internacional