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New findings on protein expression in copper deficient rat heart using proteomic approaches
Author(s) -
Zeng Huawei,
Saari Jack T.
Publication year - 2006
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.20.4.a553-a
Subject(s) - protein expression , microbiology and biotechnology , chemistry , copper , computational biology , biology , biochemistry , gene , organic chemistry
Dietary Cu‐deficiency not only causes a hypertrophic cardiomyopathy but also increases cancer risk in rodent models. The present studies were undertaken to determine the effect of Cu‐deficiency on protein profiles in rat heart tissue. Male Sprague‐Dawley rats were fed diets that were either copper adequate (6.0 μg copper/g diet n = 6) or copper deficient (0.3 μg copper/g diet n = 6) for 5 wk. An approach using liquid chromatography mass spectrometry analysis coupled with DNA gel shift and immunodection dot blotting revealed a 132 kDa protein complex in the protein extract from hearts of Cu‐deficient, but not Cu‐adequate rats. This complex contained a collagen alpha (I) chain precursor as well as a leucine‐rich protein 130 (DNA binding protein). These data suggest that Cu‐deficiency may have a potential to influence DNA stability and mRNA transcript processing. Another approach to determine the effect of Cu deficiency on protein profiles used the combination of the isotope‐coded affinity tag (ICAT) method and Western blotting analysis. ICAT analysis suggested that protein profiles from heart tissue of Cu‐deficient rats were different from those of Cu‐adequate rats; seven major protein species differed by more than a 100% increase or a 50% decrease. Western blotting analysis confirmed an 85% increase in fibulin‐5 (also known DANCE/EVEC) in Cu‐deficient rat heart, and suggested a critical biological function of Cu in controlling vascular redox state, scaffolding of cells to elastic fiber, and preventing elastinopathy in the rat heart.