Solution and interfacial self-assembly of Bacillus Subtilis bacterial lipoteichoic acid (LTA): Nanoclustering, and effects of Ca2+ and temperature

Bhaveshkumar Bharatiya; Magdalena K Wlodek; Robert L Harniman; Ralf  Schweins; Judith M Mantell; Gang Wang; Piotr Warszyński; Wuge H Briscoe

doi:10.1039/D2NR00595F

Solution and interfacial self-assembly of Bacillus Subtilis bacterial lipoteichoic acid (LTA): Nanoclustering, and effects of Ca2+ and temperature

Bhaveshkumar Bharatiya
, Magdalena K Wlodek
, Robert L Harniman
, Ralf Schweins
, Judith M Mantell
, Gang Wang
, Piotr Warszyński
, Wuge H Briscoe^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

1 Citation (Scopus)

74 Downloads (Pure)

Abstract

Lipoteichoic acid (LTA) is a major structural and functional molecule in the Gram-positive bacteria membrane. Knowledge of LTA adsorption at interfaces and its solution self-assembly is crucial to understanding its role in bacterial adhesion and colonisation, infections and inflammations. Here, we report the self-assembly behaviour of LTA extracted from Bacillus Subtilis, a Gram-positive bacterium, in an aqueous solution using cryogenic transmission electron microscopy (Cryo-TEM) and small-angle neutron scattering (SANS) and its adsorption behaviour at the solid-liquid interface using atomic force microscopy (AFM) imaging and quartz crystal microbalance with dissipation monitoring (QCM-D). The Cryo-TEM results indicated the formation of spherical LTA micelles that decreased in size on addition of calcium chloride (CaCl2), attributed to charge neutralisation and possible formation of stable Ca2+-bridges between the phosphate groups on neighbouring LTA chains. Analysis of the SANS data from the polydisperse LTA aggregates in solution using the two Lorentzian model revealed the existence of two correlation lengths, which could respectively account for the presence of LTA micelle clusters and the local structure arising from LTA intra-molecular interactions. In the presence of CaCl2, the decrease in the correlation lengths of the clusters indicated possible disruption of H-bonding by Ca2+, leading to poorer water-LTA interactions. At higher temepratures, the correlation length corresponding to the clusters increased, indicating a temperature assisted growth caused by the fluidization of micellar core and dehydration of the polar LTA chains. AFM imaging showed that adsorption of LTA aggregates at the SiO2-water interface was significantly prompted by the addition of CaCl2, also confirmed by QCM-D measurements. These unprecedented nanoscopic structural details on the morphology of LTA aggregates in solution and at the solid-liquid interface add to our fundamental understanding of its self-assembly behaviour hitherto underexplored.

Original language	English
Pages (from-to)	12265-12274
Number of pages	10
Journal	Nanoscale
Volume	14
Issue number	34
DOIs	https://doi.org/10.1039/D2NR00595F
Publication status	Published - 18 Jul 2022

Bibliographical note

Funding Information:
We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [ https://doi.ill.fr/10.5291/ILL-DATA.9-13-912 ]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS.

Funding Information:
We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [https://doi.ill.fr/10.5291/ILL-DATA.9-13-912]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS.

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© 2022 The Royal Society of Chemistry.

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title = "Solution and interfacial self-assembly of Bacillus Subtilis bacterial lipoteichoic acid (LTA): Nanoclustering, and effects of Ca2+ and temperature",

abstract = "Lipoteichoic acid (LTA) is a major structural and functional molecule in the Gram-positive bacteria membrane. Knowledge of LTA adsorption at interfaces and its solution self-assembly is crucial to understanding its role in bacterial adhesion and colonisation, infections and inflammations. Here, we report the self-assembly behaviour of LTA extracted from Bacillus Subtilis, a Gram-positive bacterium, in an aqueous solution using cryogenic transmission electron microscopy (Cryo-TEM) and small-angle neutron scattering (SANS) and its adsorption behaviour at the solid-liquid interface using atomic force microscopy (AFM) imaging and quartz crystal microbalance with dissipation monitoring (QCM-D). The Cryo-TEM results indicated the formation of spherical LTA micelles that decreased in size on addition of calcium chloride (CaCl2), attributed to charge neutralisation and possible formation of stable Ca2+-bridges between the phosphate groups on neighbouring LTA chains. Analysis of the SANS data from the polydisperse LTA aggregates in solution using the two Lorentzian model revealed the existence of two correlation lengths, which could respectively account for the presence of LTA micelle clusters and the local structure arising from LTA intra-molecular interactions. In the presence of CaCl2, the decrease in the correlation lengths of the clusters indicated possible disruption of H-bonding by Ca2+, leading to poorer water-LTA interactions. At higher temepratures, the correlation length corresponding to the clusters increased, indicating a temperature assisted growth caused by the fluidization of micellar core and dehydration of the polar LTA chains. AFM imaging showed that adsorption of LTA aggregates at the SiO2-water interface was significantly prompted by the addition of CaCl2, also confirmed by QCM-D measurements. These unprecedented nanoscopic structural details on the morphology of LTA aggregates in solution and at the solid-liquid interface add to our fundamental understanding of its self-assembly behaviour hitherto underexplored.",

author = "Bhaveshkumar Bharatiya and Wlodek, \{Magdalena K\} and Harniman, \{Robert L\} and Ralf Schweins and Mantell, \{Judith M\} and Gang Wang and Piotr Warszy{\'n}ski and Briscoe, \{Wuge H\}",

note = "Funding Information: We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [ https://doi.ill.fr/10.5291/ILL-DATA.9-13-912 ]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Sk{\l}odowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS. Funding Information: We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [https://doi.ill.fr/10.5291/ILL-DATA.9-13-912]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Sk{\l}odowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS. Publisher Copyright: {\textcopyright} 2022 The Royal Society of Chemistry.",

year = "2022",

month = jul,

day = "18",

doi = "10.1039/D2NR00595F",

language = "English",

volume = "14",

pages = "12265--12274",

journal = "Nanoscale",

issn = "2040-3364",

publisher = "Royal Society of Chemistry",

number = "34",

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TY - JOUR

T1 - Solution and interfacial self-assembly of Bacillus Subtilis bacterial lipoteichoic acid (LTA)

T2 - Nanoclustering, and effects of Ca2+ and temperature

AU - Bharatiya, Bhaveshkumar

AU - Wlodek, Magdalena K

AU - Harniman, Robert L

AU - Schweins, Ralf

AU - Mantell, Judith M

AU - Wang, Gang

AU - Warszyński, Piotr

AU - Briscoe, Wuge H

N1 - Funding Information: We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [ https://doi.ill.fr/10.5291/ILL-DATA.9-13-912 ]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS. Funding Information: We acknowledge the Institut Laue-Langevin (ILL) for the awarded beamtime under experiment number: 9-13-912 [https://doi.ill.fr/10.5291/ILL-DATA.9-13-912]. We thank Lauren Matthews and Xueying Guo for their help with the SANS measurements and useful discussions. We also acknowledge the Wolfson Foundation for establishing the Wolfson Bioimaging Facility, University of Bristol. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement SoftNanoHybrid no. 797038 (Bhavesh Bharatiya) and grant agreement HYDRA no. 793574 (Gang Wang). WHB would like to acknowledge funding from Engineering and Physical Science Research Council (EPSRC; EP/H034862/1 and EP/H014861/1). MW and PW acknowledge support from the Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS. Publisher Copyright: © 2022 The Royal Society of Chemistry.

PY - 2022/7/18

Y1 - 2022/7/18

N2 - Lipoteichoic acid (LTA) is a major structural and functional molecule in the Gram-positive bacteria membrane. Knowledge of LTA adsorption at interfaces and its solution self-assembly is crucial to understanding its role in bacterial adhesion and colonisation, infections and inflammations. Here, we report the self-assembly behaviour of LTA extracted from Bacillus Subtilis, a Gram-positive bacterium, in an aqueous solution using cryogenic transmission electron microscopy (Cryo-TEM) and small-angle neutron scattering (SANS) and its adsorption behaviour at the solid-liquid interface using atomic force microscopy (AFM) imaging and quartz crystal microbalance with dissipation monitoring (QCM-D). The Cryo-TEM results indicated the formation of spherical LTA micelles that decreased in size on addition of calcium chloride (CaCl2), attributed to charge neutralisation and possible formation of stable Ca2+-bridges between the phosphate groups on neighbouring LTA chains. Analysis of the SANS data from the polydisperse LTA aggregates in solution using the two Lorentzian model revealed the existence of two correlation lengths, which could respectively account for the presence of LTA micelle clusters and the local structure arising from LTA intra-molecular interactions. In the presence of CaCl2, the decrease in the correlation lengths of the clusters indicated possible disruption of H-bonding by Ca2+, leading to poorer water-LTA interactions. At higher temepratures, the correlation length corresponding to the clusters increased, indicating a temperature assisted growth caused by the fluidization of micellar core and dehydration of the polar LTA chains. AFM imaging showed that adsorption of LTA aggregates at the SiO2-water interface was significantly prompted by the addition of CaCl2, also confirmed by QCM-D measurements. These unprecedented nanoscopic structural details on the morphology of LTA aggregates in solution and at the solid-liquid interface add to our fundamental understanding of its self-assembly behaviour hitherto underexplored.

AB - Lipoteichoic acid (LTA) is a major structural and functional molecule in the Gram-positive bacteria membrane. Knowledge of LTA adsorption at interfaces and its solution self-assembly is crucial to understanding its role in bacterial adhesion and colonisation, infections and inflammations. Here, we report the self-assembly behaviour of LTA extracted from Bacillus Subtilis, a Gram-positive bacterium, in an aqueous solution using cryogenic transmission electron microscopy (Cryo-TEM) and small-angle neutron scattering (SANS) and its adsorption behaviour at the solid-liquid interface using atomic force microscopy (AFM) imaging and quartz crystal microbalance with dissipation monitoring (QCM-D). The Cryo-TEM results indicated the formation of spherical LTA micelles that decreased in size on addition of calcium chloride (CaCl2), attributed to charge neutralisation and possible formation of stable Ca2+-bridges between the phosphate groups on neighbouring LTA chains. Analysis of the SANS data from the polydisperse LTA aggregates in solution using the two Lorentzian model revealed the existence of two correlation lengths, which could respectively account for the presence of LTA micelle clusters and the local structure arising from LTA intra-molecular interactions. In the presence of CaCl2, the decrease in the correlation lengths of the clusters indicated possible disruption of H-bonding by Ca2+, leading to poorer water-LTA interactions. At higher temepratures, the correlation length corresponding to the clusters increased, indicating a temperature assisted growth caused by the fluidization of micellar core and dehydration of the polar LTA chains. AFM imaging showed that adsorption of LTA aggregates at the SiO2-water interface was significantly prompted by the addition of CaCl2, also confirmed by QCM-D measurements. These unprecedented nanoscopic structural details on the morphology of LTA aggregates in solution and at the solid-liquid interface add to our fundamental understanding of its self-assembly behaviour hitherto underexplored.

U2 - 10.1039/D2NR00595F

DO - 10.1039/D2NR00595F

M3 - Article (Academic Journal)

SN - 2040-3364

VL - 14

SP - 12265

EP - 12274

JO - Nanoscale

JF - Nanoscale

IS - 34

ER -

Solution and interfacial self-assembly of Bacillus Subtilis bacterial lipoteichoic acid (LTA): Nanoclustering, and effects of Ca2+ and temperature

Abstract

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