RTX Adhesins are Key Bacterial Surface Megaproteins in the Formation of Biofilms

Guo, Shuaiqi; Vance, Tyler D. R.; Stevens, Corey A.; Voets, Ilja K.; Davies, Peter L.

doi:10.1016/j.tim.2019.02.001

Cited by 17 publications

(35 citation statements)

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“…MpIBP has a Cterminal signal sequence enabling its secretion by the Type I Secretion System (T1SS) specific to M. primoryensis ( Fig.2a) [17]. At the N terminus, MpIBP contains an "anchor module" that helps retain the protein to the cell surface by plugging the T1SS outer-membrane pore [17,18,[21][22][23]. The protein further consists of 120 identical 104-aa tandem repeats that serve to project a set of ligand-binding modules away from the cell surface to interact with their target molecules, including various carbohydrates, proteins, and ice.…”

Section: Cell-surface Display Of Mpamentioning

confidence: 99%

“…The protein further consists of 120 identical 104-aa tandem repeats that serve to project a set of ligand-binding modules away from the cell surface to interact with their target molecules, including various carbohydrates, proteins, and ice. The sugar-binding domain (SBD) of MpIBP binds to glucose in a Ca 2+ -dependent manner [17,18,24], and is used in this study to allow engineered bacteria to bind to dextran-based hydrogels.…”

Section: Cell-surface Display Of Mpamentioning

confidence: 99%

“…We engineer E. coli bacteria to surface-display a Ca 2+ -dependent glucose-binding adhesin which has strong interactions with dextran-based hydrogels ( Fig.1b and 1c for details). The engineered adhesin, referred to as MpA, is derived from the Marinomonas primoryensis ice-binding protein (MpIBP) which helps these bacteria to assemble into multi-species biofilms on ice [16][17][18]. Compared to conventional living material designs, our hydrogel-based material has large pores with diameters ranging from 10-100 µM, which substantially improves the diffusion of macromolecules through the matrix and bypasses complicated fabrication procedures.…”

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Engineered Living Materials based on Adhesin-mediated Trapping of Programmable Cells

Guo¹,

Dubuc²,

Rave

et al. 2019

Preprint

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5 Stichting PAMM, Laboratory for pathology and medical microbiology, De Run 6250, 5504 DL Veldhoven 6 Laboratory of Physical Chemistry 7 Molecular biosensing for medical diagnostics 8 Laboratory of protein engineering 1-4, 6-8 : Abstract:Engineered living materials have the potential for wide-ranging applications such as biosensing and treatment of diseases. Programmable cells provide the functional basis for living materials, however, their release into the environment raises numerous biosafety concerns. Current designs that limit the release of genetically engineered cells typically involve the fabrication of multi-layer hybrid materials with sub-micron porous matrices. Nevertheless the stringent physical barriers limit the diffusion of macromolecules and therefore the repertoire of molecules available for actuation in response to communication signals between cells and their environment. Here, we engineer a first-of-its-kind living material entitled 'Platform for Adhesin-mediated Trapping of Cells in Hydrogels' (PATCH). This technology is based on engineered E. coli that displays an adhesion protein derived from an Antarctic bacterium with high affinity for glucose. The adhesin stably anchors E. coli in dextran-based hydrogels with large pore diameters (10-100 µm) and reduces the leakage of bacteria into the environment by up to 100-fold. As an application of PATCH, we engineered E. coli to secrete lysostaphin via the Type 1 Secretion System and demonstrated that living materials containing this E. coli inhibit the growth of S. aureus, including the strain resistant to methicillin (MRSA). Our tunable platform allows stable integration of programmable cells in dextran-based hydrogels without compromising free diffusion of macromolecules and could have potential applications in biotechnology and biomedicine. Introduction:Synthetic biology aims to design programmable cells that combine sensing and molecular computing operations with on-demand production of proteins that have a broad spectrum of therapeutic applications [1][2][3][4]. Engineered living materials (ELMs) integrate genetically engineered cells with free standing materials and represent a new class of environmentally responsive living devices with designer physicochemical and material properties [5][6][7]. Ideally, ELMs provide mechanical robustness to engineered cells, prevent their leakage to the environment and allow cells to be viable for extended periods of time. The containment of genetically-modified microorganisms (GMMs) within various materials has become a grand challenge for future synthetic biology applications [8]. To date, strategies for containing GMMs inside a living device are based on the physical confinement by multi-layer materials [9][10][11]. Hybrid micro-patterned devices combining layers of elastomer and microporous hydrogel enabled the exchange of information with surrounding environment via diffusion of chemical inducers and their sensing by GMMs while displaying high mechanical resilience [10]. Nevertheless, the low porosity ...

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Section: Cell-surface Display Of Mpamentioning

confidence: 99%

Section: Cell-surface Display Of Mpamentioning

confidence: 99%

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confidence: 99%

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Engineered Living Materials based on Adhesin-mediated Trapping of Programmable Cells

Guo¹,

Dubuc²,

Rave

et al. 2019

Preprint

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“…SiiE and the M. primoryensis ice-binding protein ( Mp IBP) belong to the same family of adhesion proteins, known as the RTX adhesins, which are widespread amongst Gram-negative bacteria ( Guo et al, 2019a , Guo et al, 2019b , Satchell, 2011 ). RTX adhesins have been implicated in a variety of bacterial survival strategies, including pathogen infection ( Cirillo et al, 2001 , Syed et al, 2009 , Wagner et al, 2014 ), symbiotic colonization ( Hinsa et al, 2003 , Martínez-Gil et al, 2010 ), and microbial community development ( Guo et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Essential role of calcium in extending RTX adhesins to their target

Vance

Conroy

et al. 2020

Journal of Structural Biology: X

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“…The repeats contain the common sequence structure G-G-X-G-(N/D) -D-X-(L/I/V/W/Y/F)-X (where X can be any amino acid) and are present in variable numbers ranging from six to more than 50 copies. These nonapeptide repeats form a parallel β-roll motif within a right-handed spiral, which binds Ca 2+ ions held by two sterically neighboring nonapeptide repeats [2][3][4]. Several RTX toxins have been shown to bind Ca 2+ in solution and to be dependent on Ca 2+ to exhibit their pore-forming and cytolytic activity.…”

Section: Introductionmentioning

confidence: 99%

RTX Toxins of Animal Pathogens and Their Role as Antigens in Vaccines and Diagnostics

Frey

2019

Toxins

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Exotoxins play a central role in the pathologies caused by most major bacterial animal pathogens. The large variety of vertebrate and invertebrate hosts in the animal kingdom is reflected by a large variety of bacterial pathogens and toxins. The group of repeats in the structural toxin (RTX) toxins is particularly abundant among bacterial pathogens of animals. Many of these toxins are described as hemolysins due to their capacity to lyse erythrocytes in vitro. Hemolysis by RTX toxins is due to the formation of cation-selective pores in the cell membrane and serves as an important marker for virulence in bacterial diagnostics. However, their physiologic relevant targets are leukocytes expressing β2 integrins, which act as specific receptors for RTX toxins. For various RTX toxins, the binding to the CD18 moiety of β2 integrins has been shown to be host specific, reflecting the molecular basis of the host range of RTX toxins expressed by bacterial pathogens. Due to the key role of RTX toxins in the pathogenesis of many bacteria, antibodies directed against specific RTX toxins protect against disease, hence, making RTX toxins valuable targets in vaccine research and development. Due to their specificity, several structural genes encoding for RTX toxins have proven to be essential in modern diagnostic applications in veterinary medicine.

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RTX Adhesins are Key Bacterial Surface Megaproteins in the Formation of Biofilms

Cited by 17 publications

References 0 publications

Engineered Living Materials based on Adhesin-mediated Trapping of Programmable Cells

Engineered Living Materials based on Adhesin-mediated Trapping of Programmable Cells

Essential role of calcium in extending RTX adhesins to their target

RTX Toxins of Animal Pathogens and Their Role as Antigens in Vaccines and Diagnostics

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