The high-affinity peptidoglycan binding domain of Pseudomonas phage endolysin KZ144

被引：69

作者：

Briers, Yves ^{[1
]}

Schmelcher, Mathias ^{[2
]}

Loessner, Martin J. ^{[2
]}

Hendrix, Jelle

Engelborghs, Yves ^{[3
]}

Volckaert, Guido ^{[1
,3
]}

Lavigne, Rob ^{[1
]}

机构：

[1] Katholieke Univ Leuven, Dept Biosyst, Div Gene Technol, B-3001 Heverlee, Belgium

[2] ETH, Inst Food Sci & Nutr, CH-8092 Zurich, Switzerland

[3] Katholieke Univ Leuven, Dept Chem, Lab Biomol Dynam, B-3001 Heverlee, Belgium

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2009年 / 383卷 / 02期

关键词：

Endolysin; Peptidoglycan binding domain; Affinity; Module shuffling; Chimera; BACTERIOPHAGE-PHI-KZ; WALL LYTIC ENZYME; MUREIN HYDROLASES; LYSOZYME; POLYMERASE; AERUGINOSA; EVOLUTION; GENES;

D O I：

10.1016/j.bbrc.2009.03.161

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The binding affinity of the N-terminal peptidoglycan binding domain of endolysin KZ144 (PBDKZ), originating from Pseudomonas aeruginosa bacteriophage KZ, has been examined using a fusion protein of PBDKZ and green fluorescent protein (PBDKZ-GFP). A fluorescence recovery after photobleaching analysis of bound PBDKZ-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 PBDKZ-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 (PBDKZ-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. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：187 / 191

页数：5

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