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iTRAQ proteomic analysis of extracellular matrix remodeling in aortic valve disease.

Martin-Rojas T, Mourino-Alvarez L, Alonso-Orgaz S, Rosello-Lleti E, Calvo E, Lopez-Almodovar LF, Rivera M, Padial LR, Lopez JA, de la Cuesta F, Barderas MG - Sci Rep (2015)

Bottom Line: Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms.The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses.These findings are substantiated by our previous results demonstrating differential ECM protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Physiopathology, Hospital Nacional de Parapléjicos, SESCAM, Toledo, Spain.

ABSTRACT
Degenerative aortic stenosis (AS) is the most common worldwide cause of valve replacement. The aortic valve is a thin, complex, layered connective tissue with compartmentalized extracellular matrix (ECM) produced by specialized cell types, which directs blood flow in one direction through the heart. There is evidence suggesting remodeling of such ECM during aortic stenosis development. Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms. Aortic valve samples were collected from 18 patients which underwent aortic valve replacement (50% males, mean age of 74 years) and 18 normal control valves were obtained from necropsies (40% males, mean age of 69 years). The proteome of the samples was analyzed by 2D-LC MS/MS iTRAQ methodology. The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses. These findings are substantiated by our previous results demonstrating differential ECM protein expression. The present study has demonstrated a differential ECM protein pattern in individuals with AS, therefore supporting previous evidence of a dynamic ECM remodeling in human aortic valves during AS development.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of the proteins identified in the iTRAQ experiment.Within the 712 identified proteins, 56 proteins (34 up-regulated and 19 down-regulated in stenotic aortic valves when compared to controls, in addition to 3 proteins which are increased or decreased depending on the isoform identified in stenotic valves when compared to controls) were detected in the four runs (mixtures) of the experiment carried out.
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f1: Schematic representation of the proteins identified in the iTRAQ experiment.Within the 712 identified proteins, 56 proteins (34 up-regulated and 19 down-regulated in stenotic aortic valves when compared to controls, in addition to 3 proteins which are increased or decreased depending on the isoform identified in stenotic valves when compared to controls) were detected in the four runs (mixtures) of the experiment carried out.

Mentions: To provide insight into the consequences of AS on protein expression, we compared the proteome of AS tissue with the one from healthy valves. Lysates from 4 control samples and 4 stenotic samples were iTRAQ labeled as shown in Fig. 1 of supplementary material (Figure SM1). Samples were pooled in four labeled peptide mixtures to allow the analysis of all of them in a limited number of LC-MS/MS runs avoiding labeling bias (for experimental design see supplementary material). After the labeling procedure and liquid chromatography (LC) separation, derived peptides were identified and quantified by tandem mass spectrometry (MS/MS). The data generated by liquid chromatography coupled to mass spectrometry (LC-MS/MS) analysis were searched against the human RefSeq database using Proteome Discoverer software (Thermo Scientific).


iTRAQ proteomic analysis of extracellular matrix remodeling in aortic valve disease.

Martin-Rojas T, Mourino-Alvarez L, Alonso-Orgaz S, Rosello-Lleti E, Calvo E, Lopez-Almodovar LF, Rivera M, Padial LR, Lopez JA, de la Cuesta F, Barderas MG - Sci Rep (2015)

Schematic representation of the proteins identified in the iTRAQ experiment.Within the 712 identified proteins, 56 proteins (34 up-regulated and 19 down-regulated in stenotic aortic valves when compared to controls, in addition to 3 proteins which are increased or decreased depending on the isoform identified in stenotic valves when compared to controls) were detected in the four runs (mixtures) of the experiment carried out.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664895&req=5

f1: Schematic representation of the proteins identified in the iTRAQ experiment.Within the 712 identified proteins, 56 proteins (34 up-regulated and 19 down-regulated in stenotic aortic valves when compared to controls, in addition to 3 proteins which are increased or decreased depending on the isoform identified in stenotic valves when compared to controls) were detected in the four runs (mixtures) of the experiment carried out.
Mentions: To provide insight into the consequences of AS on protein expression, we compared the proteome of AS tissue with the one from healthy valves. Lysates from 4 control samples and 4 stenotic samples were iTRAQ labeled as shown in Fig. 1 of supplementary material (Figure SM1). Samples were pooled in four labeled peptide mixtures to allow the analysis of all of them in a limited number of LC-MS/MS runs avoiding labeling bias (for experimental design see supplementary material). After the labeling procedure and liquid chromatography (LC) separation, derived peptides were identified and quantified by tandem mass spectrometry (MS/MS). The data generated by liquid chromatography coupled to mass spectrometry (LC-MS/MS) analysis were searched against the human RefSeq database using Proteome Discoverer software (Thermo Scientific).

Bottom Line: Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms.The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses.These findings are substantiated by our previous results demonstrating differential ECM protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Physiopathology, Hospital Nacional de Parapléjicos, SESCAM, Toledo, Spain.

ABSTRACT
Degenerative aortic stenosis (AS) is the most common worldwide cause of valve replacement. The aortic valve is a thin, complex, layered connective tissue with compartmentalized extracellular matrix (ECM) produced by specialized cell types, which directs blood flow in one direction through the heart. There is evidence suggesting remodeling of such ECM during aortic stenosis development. Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms. Aortic valve samples were collected from 18 patients which underwent aortic valve replacement (50% males, mean age of 74 years) and 18 normal control valves were obtained from necropsies (40% males, mean age of 69 years). The proteome of the samples was analyzed by 2D-LC MS/MS iTRAQ methodology. The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses. These findings are substantiated by our previous results demonstrating differential ECM protein expression. The present study has demonstrated a differential ECM protein pattern in individuals with AS, therefore supporting previous evidence of a dynamic ECM remodeling in human aortic valves during AS development.

No MeSH data available.


Related in: MedlinePlus