Staphylococcal biofilm-forming protein has a contiguous rod-like structure

Dominika T. Gruszka, Justyna A. Wojdyla, Richard J. Bingham, Johan P. Turkenburg, Iain W. Manfield, Annette Steward, Andrew P. Leech, Joan A. Geoghegan, Timothy J. Foster, Jane Clarke, Jennifer R. Potts

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein (S. epidermidis) promote biofilm formation through their "B" regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+-mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation- associated protein form extended fibrils on the bacterial surface. Here we report structures of E-G5 and G5-E-G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the "zinc zipper." Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.

Original languageEnglish
Pages (from-to)E1011-E1018
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number17
DOIs
Publication statusPublished - 24 Apr 2012

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Biofilms
Membrane Proteins
Staphylococcus aureus
Proteins
Protein S
Device Removal
Staphylococcus epidermidis
Infection
Thermodynamics
Zinc
Anti-Bacterial Agents
Morbidity
Equipment and Supplies
Mortality

Cite this

Gruszka, Dominika T. ; Wojdyla, Justyna A. ; Bingham, Richard J. ; Turkenburg, Johan P. ; Manfield, Iain W. ; Steward, Annette ; Leech, Andrew P. ; Geoghegan, Joan A. ; Foster, Timothy J. ; Clarke, Jane ; Potts, Jennifer R. / Staphylococcal biofilm-forming protein has a contiguous rod-like structure. In: Proceedings of the National Academy of Sciences of the United States of America. 2012 ; Vol. 109, No. 17. pp. E1011-E1018.
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abstract = "Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein (S. epidermidis) promote biofilm formation through their {"}B{"} regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+-mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation- associated protein form extended fibrils on the bacterial surface. Here we report structures of E-G5 and G5-E-G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the {"}zinc zipper.{"} Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.",
keywords = "Device infection, Protein biophysics, Protein domains, X-ray crystallography",
author = "Gruszka, {Dominika T.} and Wojdyla, {Justyna A.} and Bingham, {Richard J.} and Turkenburg, {Johan P.} and Manfield, {Iain W.} and Annette Steward and Leech, {Andrew P.} and Geoghegan, {Joan A.} and Foster, {Timothy J.} and Jane Clarke and Potts, {Jennifer R.}",
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Gruszka, DT, Wojdyla, JA, Bingham, RJ, Turkenburg, JP, Manfield, IW, Steward, A, Leech, AP, Geoghegan, JA, Foster, TJ, Clarke, J & Potts, JR 2012, 'Staphylococcal biofilm-forming protein has a contiguous rod-like structure', Proceedings of the National Academy of Sciences of the United States of America, vol. 109, no. 17, pp. E1011-E1018. https://doi.org/10.1073/pnas.1119456109

Staphylococcal biofilm-forming protein has a contiguous rod-like structure. / Gruszka, Dominika T.; Wojdyla, Justyna A.; Bingham, Richard J.; Turkenburg, Johan P.; Manfield, Iain W.; Steward, Annette; Leech, Andrew P.; Geoghegan, Joan A.; Foster, Timothy J.; Clarke, Jane; Potts, Jennifer R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, No. 17, 24.04.2012, p. E1011-E1018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Staphylococcal biofilm-forming protein has a contiguous rod-like structure

AU - Gruszka, Dominika T.

AU - Wojdyla, Justyna A.

AU - Bingham, Richard J.

AU - Turkenburg, Johan P.

AU - Manfield, Iain W.

AU - Steward, Annette

AU - Leech, Andrew P.

AU - Geoghegan, Joan A.

AU - Foster, Timothy J.

AU - Clarke, Jane

AU - Potts, Jennifer R.

PY - 2012/4/24

Y1 - 2012/4/24

N2 - Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein (S. epidermidis) promote biofilm formation through their "B" regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+-mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation- associated protein form extended fibrils on the bacterial surface. Here we report structures of E-G5 and G5-E-G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the "zinc zipper." Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.

AB - Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein (S. epidermidis) promote biofilm formation through their "B" regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+-mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation- associated protein form extended fibrils on the bacterial surface. Here we report structures of E-G5 and G5-E-G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the "zinc zipper." Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.

KW - Device infection

KW - Protein biophysics

KW - Protein domains

KW - X-ray crystallography

UR - http://www.scopus.com/inward/record.url?scp=84860149039&partnerID=8YFLogxK

U2 - 10.1073/pnas.1119456109

DO - 10.1073/pnas.1119456109

M3 - Article

VL - 109

SP - E1011-E1018

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 17

ER -