title: Quantitative measurement of collagen methylation by capillary electrophoresis.
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.
authors.affiliation: Institute of Bioengineering and Nanotechnology, National University of Singapore.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Hong-Ping
authors.initials: HP
authors.lastname: Wei
authors.forename: Chai-Hoon
authors.initials: CH
authors.lastname: Quek
authors.forename: Ser-Mien
authors.initials: SM
authors.lastname: Chia
authors.forename: Hanry
authors.initials: H
authors.lastname: Yu
mesh_terms: 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
chemical_list: D002021:Buffers; D003094:Collagen