title: Effect of alteration of translation error rate on enzyme microheterogeneity as assessed by variation in single molecule electrophoretic mobility and catalytic activity.
abstract: The role of translation error for Escherichia coli individual beta-galactosidase molecule catalytic and electrophoretic heterogeneity was investigated using CE-LIF. An E. coli rpsL mutant with a hyperaccurate translation phenotype produced enzyme molecules that exhibited significantly less catalytic heterogeneity but no reduction of electrophoretic heterogeneity. Enzyme expressed with streptomycin-induced translation error had increased thermolability, lower activity, and no significant change to catalytic or electrophoretic heterogeneity. Modeling of the electrophoretic behaviour of beta-galactosidase suggested that variation of the hydrodynamic radius may be the most significant contributor to electrophoretic heterogeneity.
authors.affiliation: Chemistry Department, University of Winnipeg, Winnipeg, Canada.
authors.forename: Ellert R
authors.initials: ER
authors.lastname: Nichols
authors.forename: Elnaz
authors.initials: E
authors.lastname: Shadabi
authors.forename: Douglas B
authors.initials: DB
authors.lastname: Craig
mesh_terms: D055162:Biocatalysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004790:Enzyme Induction; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development; D029968:Escherichia coli Proteins; D014176:Protein Biosynthesis; D000091243:Ribosomal Protein S9; D013311:Streptozocin / Q000494:pharmacology; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D029968:Escherichia coli Proteins; D000091243:Ribosomal Protein S9; C000717190:RpsI protein, E coli; D013311:Streptozocin; D001617:beta-Glucosidase