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The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N

Intrinsically disordered regions within proteins are critical elements in many biomolecular interactions and signaling pathways. Antibacterial toxins of the colicin family, which could provide new antibiotic functions against resistant bacteria, contain disordered N-terminal translocation domains (T...

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Autores principales: Johnson, Christopher L., Solovyova, Alexandra S., Hecht, Olli, Macdonald, Colin, Waller, Helen, Grossmann, J. Günter, Moore, Geoffrey R., Lakey, Jeremy H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Biophysical Society 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647543/
https://www.ncbi.nlm.nih.gov/pubmed/29045862
http://dx.doi.org/10.1016/j.bpj.2017.08.030
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author Johnson, Christopher L.
Solovyova, Alexandra S.
Hecht, Olli
Macdonald, Colin
Waller, Helen
Grossmann, J. Günter
Moore, Geoffrey R.
Lakey, Jeremy H.
author_facet Johnson, Christopher L.
Solovyova, Alexandra S.
Hecht, Olli
Macdonald, Colin
Waller, Helen
Grossmann, J. Günter
Moore, Geoffrey R.
Lakey, Jeremy H.
author_sort Johnson, Christopher L.
collection PubMed
description Intrinsically disordered regions within proteins are critical elements in many biomolecular interactions and signaling pathways. Antibacterial toxins of the colicin family, which could provide new antibiotic functions against resistant bacteria, contain disordered N-terminal translocation domains (T-domains) that are essential for receptor binding and the penetration of the Escherichia coli outer membrane. Here we investigate the conformational behavior of the T-domain of colicin N (ColN-T) to understand why such domains are widespread in toxins that target Gram-negative bacteria. Like some other intrinsically disordered proteins in the solution state of the protein, ColN-T shows dual recognition, initially interacting with other domains of the same colicin N molecule and later, during cell killing, binding to two different receptors, OmpF and TolA, in the target bacterium. ColN-T is invisible in the high-resolution x-ray model and yet accounts for 90 of the toxin’s 387 amino acid residues. To reveal its solution structure that underlies such a dynamic and complex system, we carried out mutagenic, biochemical, hydrodynamic and structural studies using analytical ultracentrifugation, NMR, and small-angle x-ray scattering on full-length ColN and its fragments. The structure was accurately modeled from small-angle x-ray scattering data by treating ColN as a flexible system, namely by the ensemble optimization method, which enables a distribution of conformations to be included in the final model. The results reveal, to our knowledge, for the first time the dynamic structure of a colicin T-domain. ColN-T is in dynamic equilibrium between a compact form, showing specific self-recognition and resistance to proteolysis, and an extended form, which most likely allows for effective receptor binding.
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spelling pubmed-56475432017-11-23 The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N Johnson, Christopher L. Solovyova, Alexandra S. Hecht, Olli Macdonald, Colin Waller, Helen Grossmann, J. Günter Moore, Geoffrey R. Lakey, Jeremy H. Biophys J Proteins Intrinsically disordered regions within proteins are critical elements in many biomolecular interactions and signaling pathways. Antibacterial toxins of the colicin family, which could provide new antibiotic functions against resistant bacteria, contain disordered N-terminal translocation domains (T-domains) that are essential for receptor binding and the penetration of the Escherichia coli outer membrane. Here we investigate the conformational behavior of the T-domain of colicin N (ColN-T) to understand why such domains are widespread in toxins that target Gram-negative bacteria. Like some other intrinsically disordered proteins in the solution state of the protein, ColN-T shows dual recognition, initially interacting with other domains of the same colicin N molecule and later, during cell killing, binding to two different receptors, OmpF and TolA, in the target bacterium. ColN-T is invisible in the high-resolution x-ray model and yet accounts for 90 of the toxin’s 387 amino acid residues. To reveal its solution structure that underlies such a dynamic and complex system, we carried out mutagenic, biochemical, hydrodynamic and structural studies using analytical ultracentrifugation, NMR, and small-angle x-ray scattering on full-length ColN and its fragments. The structure was accurately modeled from small-angle x-ray scattering data by treating ColN as a flexible system, namely by the ensemble optimization method, which enables a distribution of conformations to be included in the final model. The results reveal, to our knowledge, for the first time the dynamic structure of a colicin T-domain. ColN-T is in dynamic equilibrium between a compact form, showing specific self-recognition and resistance to proteolysis, and an extended form, which most likely allows for effective receptor binding. The Biophysical Society 2017-10-17 2017-10-17 /pmc/articles/PMC5647543/ /pubmed/29045862 http://dx.doi.org/10.1016/j.bpj.2017.08.030 Text en © 2017 Biophysical Society. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Proteins
Johnson, Christopher L.
Solovyova, Alexandra S.
Hecht, Olli
Macdonald, Colin
Waller, Helen
Grossmann, J. Günter
Moore, Geoffrey R.
Lakey, Jeremy H.
The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title_full The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title_fullStr The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title_full_unstemmed The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title_short The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N
title_sort two-state prehensile tail of the antibacterial toxin colicin n
topic Proteins
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647543/
https://www.ncbi.nlm.nih.gov/pubmed/29045862
http://dx.doi.org/10.1016/j.bpj.2017.08.030
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