Intercepting second-messenger signaling by rationally designed peptides sequestering c-di-GMP.
Proc Natl Acad Sci U S A
; 117(29): 17211-17220, 2020 07 21.
Article
en En
| MEDLINE
| ID: mdl-32611811
ABSTRACT
The bacterial second messenger cyclic diguanylate (c-di-GMP) regulates a wide range of cellular functions from biofilm formation to growth and survival. Targeting a second-messenger network is challenging because the system involves a multitude of components with often overlapping functions. Here, we present a strategy to intercept c-di-GMP signaling pathways by directly targeting the second messenger. For this, we developed a c-di-GMP-sequestering peptide (CSP) that was derived from a CheY-like c-di-GMP effector protein. CSP binds c-di-GMP with submicromolar affinity. The elucidation of the CSPâ
c-di-GMP complex structure by NMR identified a linear c-di-GMP-binding motif, in which a self-intercalated c-di-GMP dimer is tightly bound by a network of H bonds and π-stacking interactions involving arginine and aromatic residues. Structure-based mutagenesis yielded a variant with considerably higher, low-nanomolar affinity, which subsequently was shortened to 19 residues with almost uncompromised affinity. We demonstrate that endogenously expressed CSP intercepts c-di-GMP signaling and effectively inhibits biofilm formation in Pseudomonas aeruginosa, the most widely used model for serious biofilm-associated medical implications.
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Banco de datos:
MEDLINE
Asunto principal:
Péptidos
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Proteínas Bacterianas
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Sistemas de Mensajero Secundario
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Transducción de Señal
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GMP Cíclico
Idioma:
En
Año:
2020
Tipo del documento:
Article