title: Separation of tryptic peptides of native and glycated BSA using open-tubular CEC with salophene-lanthanide-Zn(2+) complex as stationary phase.
abstract: Open-tubular CEC (OT-CEC) with a new stationary phase, salophene-lanthanide-Zn(2+) complex, has been applied to the separation of tryptic peptides of native BSA and BSA glycated by glucose and ribose. Glycation of proteins (non-enzymatic modification by sugars) significantly affects their properties and it is of great importance from a physiological point of view. Separation of tryptic peptides of glycated BSA by CZE was poor because of their strong adsorption to the bare fused silica capillary. An improved separation of tryptic peptides of both native and glycated BSA was achieved by OT-CEC in the fused silica capillary non-covalently coated with salophene-lanthanide-Zn(2+) complex, which suppressed the adsorption of peptides to the capillary and via specific interactions with some (glyco)peptides enhanced selectivity of the separation. Significant differences have been found in OT-CEC analyses of tryptic hydrolysates of native and glycated BSA. In OT-CEC-UV profile of tryptic peptides of native BSA, 44 peaks could be resolved, whereas a reduced number of 38 peaks were observed in the profile of tryptic peptides of glucose-glycated BSA and only 30 peaks were found in the case of ribose-glycated BSA. The developed OT-CEC can be potentially used for monitoring of protein glycation.
authors.affiliation: Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Sedláková
authors.forename: Adam
authors.initials: A
authors.lastname: Eckhardt
authors.forename: Katerina
authors.initials: K
authors.lastname: Lacinová
authors.forename: Statis
authors.initials: S
authors.lastname: Pataridis
authors.forename: Ivan
authors.initials: I
authors.lastname: Miksík
authors.forename: Vladimír
authors.initials: V
authors.lastname: Král
authors.forename: Václav
authors.initials: V
authors.lastname: Kasicka
mesh_terms: D000327:Adsorption; D019075:Electrophoresis, Capillary / Q000379:methods*; D005947:Glucose / Q000737:chemistry*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D012459:Salicylates / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D014357:Trypsin / Q000737:chemistry*; D015032:Zinc / Q000737:chemistry*
chemical_list: D028581:Lanthanoid Series Elements; D010446:Peptide Fragments; D012459:Salicylates; C090416:salophen; D012710:Serum Albumin, Bovine; D014357:Trypsin; D005947:Glucose; D015032:Zinc