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High-performance liquid chromatography-based assay for the quantitation of protein maleylation at the picomole level
M N McCall1, S B Easterbrook-Smith
1Department of Biochemistry, University of Sydney, New South Wales, Australia.
Analytical Biochemistry
|November 1, 1987
Abstract:
A new HPLC-based assay for the direct determination of the extent of maleylation of proteins is described. The rationale is founded on the HPLC detection of maleic acid that results from cleavage of maleyl lysyls by low pH treatment. The method, employing a C-18 reverse-phase column eluted isocratically with 0.01 M acetic acid, pH 5.4, and absorbance monitored at 210 nm, is sensitive (lower limit 1 pmol) and linear in the range 1 pmol-0.5 mumol and allows recovery of the native protein.