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Quantitative measurement of collagen methylation by capillary electrophoresis

✍ Scribed by Jing Zhang; Hong-Ping Wei; Chai-Hoon Quek; Ser-Mien Chia; Hanry Yu

Publisher
John Wiley and Sons
Year
2004
Tongue
English
Weight
115 KB
Volume
25
Category
Article
ISSN
0173-0835

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✦ Synopsis

Abstract

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 mM, 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.

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