Moenomycin-mediated affinity purification of penicillin-binding protein 1b.
The antibiotic moenomycin A inhibits the biosynthesis of peptidoglycan, the main structural polymer of the bacterial cell wall. The inhibition is based on a reversible binding of the antibiotic to one of the substrate binding sites at enzymes such as the penicillin binding protein 1b (PBP 1b). This binding has been employed to isolate PBP 1b by affinity chromatography. Suitable ligands have been prepared from moenomycin A and coupled both to affinity supports and to surface plasmon resonance sensor surfaces. The reactions that take place upon immobilization of the ligands to the affinity support and the sensor surface, respectively, have been studied in detail. With the help of surface plasmon resonance the optimal conditions for binding of PBP 1b to moenomycin-derivated ligands have been established. For the first time the selective binding of the moenomycin sugar moiety to the enzyme has been demonstrated.
Stembera
Katherina
K
Institut für Organische Chemie, Universität Leipzig, Johannisallee 29, 04103 Leipzig, Germany.
Buchynskyy
Andrij
A
Vogel
Stefan
S
Knoll
Dietmar
D
Osman
Awad A
AA
Ayala
Juan A
JA
Welzel
Peter
P
D001426:Bacterial Proteins; D001455:Bambermycins / Q000737:chemistry* / Q000494:pharmacology; D002352:Carrier Proteins; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000737:chemistry*; D006602:Hexosyltransferases / Q000737:chemistry / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000737:chemistry; D009097:Multienzyme Complexes / Q000737:chemistry / Q000302:isolation & purification*; D002267:Muramoylpentapeptide Carboxypeptidase; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases / Q000737:chemistry / Q000302:isolation & purification*; D011485:Protein Binding
D001426:Bacterial Proteins; D002352:Carrier Proteins; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D046915:Penicillin-Binding Proteins; D001455:Bambermycins; C437243:moenomycin A; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase