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University of Nebraska - Lincoln

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

2005

Biotin ? cell signaling ? HepG2 cells ? human ? tyrosine kinase

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High-Throughput Immunoblotting Identifies Biotin-Dependent Signaling Proteins In Hepg2 Hepatocarcinoma Cells1, Rocio Rodriguez-Melendez, Jacob B. Griffin, Gautam Sarath, Janos Zempleni Jun 2005

High-Throughput Immunoblotting Identifies Biotin-Dependent Signaling Proteins In Hepg2 Hepatocarcinoma Cells1, Rocio Rodriguez-Melendez, Jacob B. Griffin, Gautam Sarath, Janos Zempleni

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Biotin affects the abundance of mRNA coding for ~10% of genes expressed in human-derived hepatocarcinoma (HepG2) cells. Here, we determined whether effects of biotin on gene expression are associated with changes in the abundance of distinct proteins in cell signaling and structure. HepG2 cells were cultured in media containing the following concentrations of biotin: 0.025 nmol/L (denoted “deficient”), 0.25 nmol/L (“physiological” = control), and 10 nmol/L (“pharmacological”) for 10 d before harvesting. The abundance of 1009 proteins from whole-cell extracts was quantified by using high-throughput immunoblots. The abundance of 44 proteins changed by at least 25% in biotin-deficient and biotin-supplemented …