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Separation and identification of four distinct serine‐phosphorylation states of ovalbumin by P hos‐tag affinity electrophoresis
Author(s) -
KinoshitaKikuta Emiko,
Kinoshita Eiji,
Koike Tohru
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100518
Subject(s) - ovalbumin , polyacrylamide gel electrophoresis , phosphorylation , serine , gel electrophoresis , phosphate , sodium dodecyl sulfate , microbiology and biotechnology , biochemistry , chemistry , biology , chromatography , antigen , enzyme , immunology
Ovalbumin ( OVA ) derived from egg white contains two residues that can be phosphorylated: S er‐68 and S er‐344. Native polyacrylamide gel electrophoresis (PAGE) shows the presence of three distinct migration bands corresponding to phosphorylation states with two, one, or no phosphate groups, respectively. Phosphate‐affinity sodium dodecyl sulfate ‐ polyacrylamide gel electrophoresis ( SD S‐ PAGE ; Z n 2+ – P hos‐tag SDS ‐ PAGE ), on the other hand, showed the presence of four distinct phosphorylated states in intact OVA . In addition to the diphosphorylated and nonphosphorylated forms, two distinct species, one with a phosphate group at S er‐68 and one with a phosphate group at S er‐344, were separately visualized. The content of the OVA monophosphorylated at S er‐68 was greater than that of OVA monophosphorylated at S er‐344. Z n 2+ – P hos‐tag SDS ‐ PAGE is therefore a useful method for the quantitative analysis of the detailed phosphorylation status of food proteins.