High-performance liquid chromatography-based assay for the quantitation of protein maleylation at the picomole level.
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.
McCall
M N
MN
Department of Biochemistry, University of Sydney, New South Wales, Australia.
Easterbrook-Smith
S B
SB
D002851:Chromatography, High Pressure Liquid; D005663:Furans; D006639:Histidine; D008299:Maleic Anhydrides; D011506:Proteins / Q000032:analysis*; D014302:Trinitrobenzenesulfonic Acid; D014462:Ultrafiltration
D005663:Furans; D008299:Maleic Anhydrides; D011506:Proteins; D006639:Histidine; D014302:Trinitrobenzenesulfonic Acid