Title: The high-affinity peptidoglycan binding domain of Pseudomonas phage endolysin KZ144
Authors: Briers, Yves
Schmelcher, Mathias
Loessner, Martin J
Hendrix, Jelle
Engelborghs, Yves
Volckaert, Guido
Lavigne, Rob # ×
Issue Date: May-2009
Publisher: Academic Press
Series Title: Biochemical and Biophysical Research Communications vol:383 issue:2 pages:187-191
Abstract: The binding affinity of the N-terminal peptidoglycan binding domain of endolysin KZ144 (PBD(KZ)), originating from Pseudomonas aeruginosa bacteriophage varphiKZ, has been examined using a fusion protein of PBD(KZ) and green fluorescent protein (PBD(KZ)-GFP). A fluorescence recovery after photobleaching analysis of bound PBD(KZ)-GFP molecules showed less than 10% fluorescence recovery in the bleached area within 15 min. Surface plasmon resonance analysis confirmed this apparent high binding affinity revealing an equilibrium affinity constant of 2.95 x 10(7)M(-1) for the PBD(KZ)-peptidoglycan interaction. This unique domain, which binds to the peptidoglycan of all tested Gram-negative species, was harnessed to improve the specific activity of the peptidoglycan hydrolase domain KMV36C. The chimeric peptidoglycan hydrolase (PBD(KZ)-KMV36C) exhibits a threefold higher specific activity than the native catalytic domain (KMV36C). These results demonstrate that the modular assembly of functional domains is a rational approach to improve the specific activity of endolysins from phages infecting Gram-negatives.
ISSN: 0006-291X
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Division of Gene Technology (-)
Biochemistry, Molecular and Structural Biology Section
× corresponding author
# (joint) last author

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