Entropically Driven Self‐Assembly of <i>Lysinibacillus sphaericus</i> S‐Layer Proteins Analyzed Under Various Environmental Conditions

Teixeira, Leonardo Maestri; Strickland, Aaron D.; Mark, Sonny S.; Bergkvist, Magnus; Sierra‐Sastre, Yajaira; Batt, Carl A.

doi:10.1002/mabi.200900175

Cited by 17 publications

(19 citation statements)

References 54 publications

(53 reference statements)

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“…A detailed discussion of the specific role of Ca 2+ -ions (and other divalent cations) associated with conformational changes within a protein and a more generic role mediating interactions between proteins (and supports) may be found in references [27,28,29]. The reassembly is entropy-driven and a fascinating example of matrix assembly following a multistage, non-classical pathway in which the process of S-layer protein folding is directly linked with assembly into extended clusters [13]. In a theoretical approach, the formation of an S-layer lattice with square (p4) lattice symmetry (as for SbpA) was investigated by assuming both non-specific attractive and specific directional bonds for the monomers [30,31,32].…”

Section: Resultsmentioning

confidence: 99%

“…In vitro self-assembly studies in solutions showed that self-assembly products (flat sheets, cylinders) are formed during dialysis of the disrupting agent against selected buffer solutions (ionic strength and pH) [2]. Kinetic measurements at different temperatures indicated that the assembly process is entropy driven with a rapid initial phase, at which oligomeric precursor patches acting as nucleation sites are formed, and a consecutive slow phase of crystal growth [12,13]. …”

Section: Introductionmentioning

confidence: 99%

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In Vitro Characterization of the Two-Stage Non-Classical Reassembly Pathway of S-Layers

Breitwieser

Iturri

Toca-Herrera

et al. 2017

IJMS

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The recombinant bacterial surface layer (S-layer) protein rSbpA of Lysinibacillus sphaericus CCM 2177 is an ideal model system to study non-classical nucleation and growth of protein crystals at surfaces since the recrystallization process may be separated into two distinct steps: (i) adsorption of S-layer protein monomers on silicon surfaces is completed within 5 min and the amount of bound S-layer protein sufficient for the subsequent formation of a closed crystalline monolayer; (ii) the recrystallization process is triggered—after washing away the unbound S-layer protein—by the addition of a CaCl2 containing buffer solution, and completed after approximately 2 h. The entire self-assembly process including the formation of amorphous clusters, the subsequent transformation into crystalline monomolecular arrays, and finally crystal growth into extended lattices was investigated by quartz crystal microbalance with dissipation (QCM-D) and atomic force microscopy (AFM). Moreover, contact angle measurements showed that the surface properties of S-layers change from hydrophilic to hydrophobic as the crystallization proceeds. This two-step approach is new in basic and application driven S-layer research and, most likely, will have advantages for functionalizing surfaces (e.g., by spray-coating) with tailor-made biological sensing layers.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In Vitro Characterization of the Two-Stage Non-Classical Reassembly Pathway of S-Layers

Breitwieser

Iturri

Toca-Herrera

et al. 2017

IJMS

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“…3,4 Due to its excellent ability to regularly assemble monomers in aqueous environments, Slp can spontaneously self-assemble on air-water interfaces, solid supports, or liposomes or in suspension and solution, by an entropydriven process (Figure 1). 5,6 This feature enables a wide range of potential applications of Slp in nanomedicine, bioactive drug delivery, and gene drug delivery. As a result, Slp-coated liposomes have received increasing attention in the past decade.…”

Section: Introductionmentioning

confidence: 99%

<p>Slp-coated liposomes for drug delivery and biomedical applications: potential and challenges</p>

Luo¹,

Yang²,

Yao³

et al. 2019

IJN

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Slp forms a crystalline array of proteins on the outermost envelope of bacteria and archaea with a molecular weight of 40-200 kDa. Slp can self-assemble on the surface of liposomes in a proper environment via electrostatic interactions, which could be employed to functionalize liposomes by forming Slp-coated liposomes for various applications. Among the molecular characteristics, the stability, adhesion, and immobilization of biomacromolecules are regarded as the most meaningful. Compared to plain liposomes, Slp-coated liposomes show excellent physicochemical and biological stabilities. Recently, Slp-coated liposomes were shown to specifically adhere to the gastrointestinal tract, which was attributed to the "ligand-receptor interaction" effect. Furthermore, Slp as a "bridge" can immobilize functional biomacromolecules on the surface of liposomes via protein fusion technology or intermolecular forces, endowing liposomes with beneficial functions. In view of these favorable features, Slp-coated liposomes are highly likely to be an ideal platform for drug delivery and biomedical uses. This review aims to provide a general framework for the structure and characteristics of Slp and the interactions between Slp and liposomes, to highlight the unique properties and drug delivery as well as the biomedical applications of the Slp-coated liposomes, and to discuss the ongoing challenges and perspectives.

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“…To produce SbpA under non-denaturing conditions, we amplified the SbpA gene from the genomic DNA of Lysinibacillus sphaericus (ATCC number 4525) [7, 9], cloned it into the pET28a vector following an established protocol [9] and expressed the protein in E. coli strain HMS174(DE3) (EMD Biosciences). Once induced, cells were cultured for 18 hours at 18°C, and then harvested and lysed as described before [15].…”

Section: Resultsmentioning

confidence: 99%

Fast and easy protocol for the purification of recombinant S‐layer protein for synthetic biology applications

Norville

Kelly

Knight³

et al. 2011

Biotechnology Journal

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A goal of synthetic biology is to make biological systems easier to engineer. One of the aims is to design – with nanometer-scale precision – biomaterials with well-defined properties. The surface layer protein SbpA forms two-dimensional (2D) arrays naturally on the cell surface of Lysinibacillus sphaericus but also as purified protein in solution upon addition of divalent cations. Its high propensity to form crystalline arrays, the simple way by which its crystallization can be controlled by divalent cations and the possibility to genetically alter the protein make SbpA an attractive molecule for synthetic biology. To be a useful tool, however, it is important that a simple protocol can be used to produce recombinant wild-type as well as modified SbpA in large quantities and in a biologically active form. The present study addresses this requirement by introducing a mild and non-denaturing purification protocol to produce milligram quantities of recombinant, active SbpA.

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Entropically Driven Self‐Assembly of Lysinibacillus sphaericus S‐Layer Proteins Analyzed Under Various Environmental Conditions

Cited by 17 publications

References 54 publications

In Vitro Characterization of the Two-Stage Non-Classical Reassembly Pathway of S-Layers

In Vitro Characterization of the Two-Stage Non-Classical Reassembly Pathway of S-Layers

<p>Slp-coated liposomes for drug delivery and biomedical applications: potential and challenges</p>

Fast and easy protocol for the purification of recombinant S‐layer protein for synthetic biology applications

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