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Quantitative measurement of collagen methylation by capillary electrophoresis
Author(s) -
Zhang Jing,
Wei HongPing,
Quek ChaiHoon,
Chia SerMien,
Yu Hanry
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406071
Subject(s) - methylation , capillary electrophoresis , chemistry , phosphate buffered saline , chromatography , polyvinyl alcohol , capillary action , dna methylation , electrophoresis , biophysics , biochemistry , dna , materials science , biology , gene expression , organic chemistry , composite material , gene
Collagen methylation has been exploited in various applications involving living cells. We have observed correlation between the collagen methylation with the rate of cell proliferation in three‐dimensional (3‐D) microenvironment. To quantify the degree of collagen methylation, we have developed a capillary zone electrophoresis method. Using a polyvinyl alcohol‐coated fused‐silica capillary and UV detection at 200 nm, we have optimized pH and separated the native collagen into three major bands in phosphate buffer (50 m M , pH 2.5) with 0.05% hydroxypropylmethylcellulose. Under these conditions, the methylated collagens were separated into four major bands, which changed with different methylation reaction conditions. We propose an index to quantify the degree of collagen methylation that also correlates with their effects on cell proliferation.