Quantitative measurement of collagen methylation by capillary electrophoresis.
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.
Zhang
Jing
J
Institute of Bioengineering and Nanotechnology, National University of Singapore.
Wei
Hong-Ping
HP
Quek
Chai-Hoon
CH
Chia
Ser-Mien
SM
Yu
Hanry
H
D000818:Animals; D002021:Buffers; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003094:Collagen / Q000032:analysis* / Q000378:metabolism; D019075:Electrophoresis, Capillary; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008745:Methylation; D011499:Protein Processing, Post-Translational; D051381:Rats
D002021:Buffers; D003094:Collagen