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Proteomic validation: Searching for a heart of gold
Author(s) -
Parsons Hannah L,
Kulpa Jerzy,
Brownsey Roger W.,
Wambolt Richard W,
Allard Michael F.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1162.3
Subject(s) - vdac1 , isobaric labeling , proteome , proteomics , chemistry , tandem mass tag , quantitative proteomics , computational biology , biochemistry , biology , escherichia coli , bacterial outer membrane , gene
Recent developments in proteomics techniques allow sensitive identification and relative quantitation of proteins in tissues and with increased sensitivity come the ability to detect small changes in protein expression. However, the means by which changes are validated remains incompletely defined. The proteome of mitochondria of hypertrophied hearts from rats with an abdominal aortic constriction (H) was quantitated using amine‐reactive isobaric tagging reagents (iTRAQ®) and tandem mass spectrometry. A small number (15 out of 250) were significantly increased and none decreased significantly. Increases ranged from 10 to 20%, as exemplified by voltage‐dependent anion channel‐1 (VDAC1), with only monoamine oxidase‐A (MAO‐A) showing a substantially greater increase (see table). Traditional immunoblot analysis revealed the significant increase in MAO‐A but not that of VDAC1. The discrepancy in results highlights the relative insensitivity of traditional immunoblot analysis and indicates that more sensitive approaches are essential. Supported by a grant from the Canadian Institutes for Health Research.