Rapid screening of a large number of immobilized textile dyes for the purification of proteins: use of penicillin-binding protein 4 of Escherichia coli as a model enzyme.
A rapid method for screening the affinity of proteins to dye-modified resins is described. Performing the binding and elution of the protein extracts in a batch-wise manner and eluting the bound proteins with SDS-PAGE denaturation buffer speed up the screening process and allow the analysis of large collections of dyes. Penicillin-binding protein 4 of Escherichia coli was used as a model enzyme to determine the influences of pH, metal ions, and ionic strength (0 to 500 mM NaCl) on its binding behavior using a collection of 98 dye-affinity resins.
Mottl
H
H
Department of Biochemistry, BIOSON Research Institute, University of Groningen, The Netherlands.
Keck
W
W
D001426:Bacterial Proteins / Q000187:drug effects / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000187:drug effects / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins; D006602:Hexosyltransferases; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D002267:Muramoylpentapeptide Carboxypeptidase / Q000187:drug effects / Q000302:isolation & purification*; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D011485:Protein Binding / Q000187:drug effects; D012492:Salts / Q000494:pharmacology; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; D014227:Triazines; D015032:Zinc / Q000494:pharmacology
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004396:Coloring Agents; D029968:Escherichia coli Proteins; D046915:Penicillin-Binding Proteins; D012492:Salts; D014227:Triazines; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; C478706:penicillin-binding protein 4, E coli; D002267:Muramoylpentapeptide Carboxypeptidase; D015032:Zinc