Protein crystal based materials for nanoscale applications in medicine and biotechnology

Hartje, Luke F.; Snow, Christopher D.

doi:10.1002/wnan.1547

Cited by 23 publications

(25 citation statements)

References 174 publications

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“…Protein crystals, especially of enzymes, have gained interest, for example, for use as 'micro-bioreactors' that serve to concentrate reactants in confined volumes, in line with other protein-encapsulation methods (Minten et al, 2009). Crystals used as bioreactors are typically not of diffraction quality, as they are stabilized to withstand harsh reaction conditions (Yan et al, 2015;Hartje & Snow, 2018), for example by glutaraldehyde cross-linking (Andersen et al, 2009). Protein crystals or their solvent channels have also been investigated as templates for nanomaterials or as drug-delivery systems (reviewed by Abe & Ueno, 2015).…”

Section: Potential Of Hostal Systems For Biotechnological Applicationsmentioning

confidence: 99%

Guest-protein incorporation into solvent channels of a protein host crystal (hostal)

Sprenger¹,

Carey²,

Schulz³

et al. 2021

Acta Cryst Sect D Struct Biol

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Soaking small molecules into the solvent channels of protein crystals is the most common method of obtaining crystalline complexes with ligands such as substrates or inhibitors. The solvent channels of some protein crystals are large enough to allow the incorporation of macromolecules, but soaking of protein guests into protein crystals has not been reported. Such protein host crystals (here given the name hostals) incorporating guest proteins may be useful for a wide range of applications in biotechnology, for example as cargo systems or for diffraction studies analogous to the crystal sponge method. The present study takes advantage of crystals of the Escherichia coli tryptophan repressor protein (ds-TrpR) that are extensively domain-swapped and suitable for incorporating guest proteins by diffusion, as they are robust and have large solvent channels. Confocal fluorescence microscopy is used to follow the migration of cytochrome c and fluorophore-labeled calmodulin into the solvent channels of ds-TrpR crystals. The guest proteins become uniformly distributed in the crystal within weeks and enriched within the solvent channels. X-ray diffraction studies on host crystals with high concentrations of incorporated guests demonstrate that diffraction limits of ∼2.5 Å can still be achieved. Weak electron density is observed in the solvent channels, but the guest-protein structures could not be determined by conventional crystallographic methods. Additional approaches that increase the ordering of guests in the host crystal are discussed that may support protein structure determination using the hostal system in the future. This host system may also be useful for biotechnological applications where crystallographic order of the guest is not required.

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Section: Potential Of Hostal Systems For Biotechnological Applicationsmentioning

confidence: 99%

Guest-protein incorporation into solvent channels of a protein host crystal (hostal)

Sprenger¹,

Carey²,

Schulz³

et al. 2021

Acta Cryst Sect D Struct Biol

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“…Protein crystals have gained interest in a variety of research domains as their properties, especially their high intrinsic order and porosity, could allow stabilizing functional proteins within the protein crystal scaffold for application in biotechnology and medicine [ 6 ]. The possibility of using protein crystals as a polyvalent nanomaterial able to concentrate, stabilize, and protect functional proteins has been envisaged for a large range of applications, notably including biosensing (i.e., detection of relevant molecules by the use of biological macromolecules), biotemplating (i.e., assembly of inorganic nanostructures guided by the protein crystal scaffold), catalysis, and vaccine/drug delivery [ 3 , 4 , 5 , 6 ].…”

Section: Functionalizing Toxin Crystals For the Development Of New Biotechnological Toolsmentioning

confidence: 99%

“…Additionnally, crystals represent a means to provide molecules at high concentration and with properties that can be tailored to provide long-term storage, controlled release, and retained activity. This is of particular interest for drug development and delivery, notably of pharmaceuticals [ 3 , 4 ], but also for catalysts that can be formulated for large-scale industrial applications [ 5 , 6 ]. Crystals therefore hold the promise of multiple applications, from the most fundamental academic research purposes to the development of innovative biotechnological products [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

“…This is of particular interest for drug development and delivery, notably of pharmaceuticals [ 3 , 4 ], but also for catalysts that can be formulated for large-scale industrial applications [ 5 , 6 ]. Crystals therefore hold the promise of multiple applications, from the most fundamental academic research purposes to the development of innovative biotechnological products [ 6 ]. However, the crystallization of macromolecules, and notably proteins, is a process hardly predictable due to the many parameters affecting the nucleation and growth of crystals [ 7 , 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

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Can (We Make) Bacillus thuringiensis Crystallize More Than Its Toxins?

Tetreau

Андреева

Banneville

et al. 2021

Toxins

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The development of finely tuned and reliable crystallization processes to obtain crystalline formulations of proteins has received growing interest from different scientific fields, including toxinology and structural biology, as well as from industry, notably for biotechnological and medical applications. As a natural crystal-making bacterium, Bacillus thuringiensis (Bt) has evolved through millions of years to produce hundreds of highly structurally diverse pesticidal proteins as micrometer-sized crystals. The long-term stability of Bt protein crystals in aqueous environments and their specific and controlled dissolution are characteristics that are particularly sought after. In this article, I explore whether the crystallization machinery of Bt can be hijacked as a means to produce (micro)crystalline formulations of proteins for three different applications: (i) to develop new bioinsecticidal formulations based on rationally improved crystalline toxins, (ii) to functionalize crystals with specific characteristics for biotechnological and medical applications, and (iii) to produce microcrystals of custom proteins for structural biology. By developing the needs of these different fields to figure out if and how Bt could meet each specific requirement, I discuss the already published and/or patented attempts and provide guidelines for future investigations in some underexplored yet promising domains.

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“…The very large solvent channels typical of ds-TrpRs are being analyzed for possible use as a host system to facilitate diffraction analysis of guest proteins that cannot be crystallized (a so-called hostal system; Sprenger et al, 2021). This system potentially offers a novel approach to the incorporation of biomolecular guests into porous systems, an area of general interest in applied biotechnology today (Yan et al, 2015;Abe & Ueno, 2015;Hartje & Snow, 2018). Successful diffraction analysis of guest proteins in any host system requires uniform guest orientation to achieve crystallographic occupancy.…”

Section: Introductionmentioning

confidence: 99%

Crystal structures of Val58Ile tryptophan repressor in a domain-swapped array in the presence and absence ofL-tryptophan

Sprenger¹,

Lawson²,

Wachenfeldt³

et al. 2021

Acta Cryst Sect F

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The crystal structures of domain-swapped tryptophan repressor (TrpR) variant Val58Ile before and after soaking with the physiological ligand L-tryptophan (L-Trp) indicate that L-Trp occupies the same location in the domain-swapped form as in native dimeric TrpR and makes equivalent residue contacts. This result is unexpected because the ligand binding-site residues arise from three separate polypeptide chains in the domain-swapped form. This work represents the first published structure of a domain-swapped form of TrpR with L-Trp bound. The presented structures also show that the protein amino-terminus, whether or not it bears a disordered extension of about 20 residues, is accessible in the large solvent channels of the domain-swapped crystal form, as in the structures reported previously in this form for TrpR without N-terminal extensions. These findings inspire the exploration of L-Trp analogs and N-terminal modifications as labels to orient guest proteins that cannot otherwise be crystallized in the solvent channels of crystalline domain-swapped TrpR hosts for potential diffraction analysis.

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Protein crystal based materials for nanoscale applications in medicine and biotechnology

Cited by 23 publications

References 174 publications

Guest-protein incorporation into solvent channels of a protein host crystal (hostal)

Guest-protein incorporation into solvent channels of a protein host crystal (hostal)

Can (We Make) Bacillus thuringiensis Crystallize More Than Its Toxins?

Crystal structures of Val58Ile tryptophan repressor in a domain-swapped array in the presence and absence ofL-tryptophan

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