Programmed Assembly of Host–Guest Protein Crystals

Huber, Thaddaus R.; Hartje, Luke F.; McPherson, Eli C.; Kowalski, Ann E.; Snow, Christopher D.

doi:10.1002/smll.201602703

Cited by 24 publications

(23 citation statements)

References 13 publications

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“…However, supplying aldehydes via gentle vapor diffusion [9] can improve outcomes. We have observed that using glyoxal and EDC can likewise result in negligible diffraction loss, particularly if the reactive chemistry is quenched [11,12]. In the case of CC1, we have once again found that carefully optimized crosslinking protocols can maintain diffraction.…”

Section: Discussionmentioning

confidence: 63%

“…In traditional protein X-ray crystallography, glutaraldehyde, a highly reactive crosslinker, can increase crystal stability in varying solution conditions, and can even improve diffraction resolution [9,10]. In our previous work on protein crystals, we have found that glyoxal offers an effective alternative to glutaraldehyde [11,12]. Chemical crosslinking and photo-crosslinking methods for DNA crystals are also established in the literature [13][14][15]; however, we wanted to focus on a protocol in which the crosslinking does not require a specific sequence of DNA and does not add atoms to the structure (a zero-length crosslink).…”

Section: Introductionmentioning

confidence: 99%

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Stabilizing DNA–Protein Co-Crystals via Intra-Crystal Chemical Ligation of the DNA

et al. 2021

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Protein and DNA co-crystals are most commonly prepared to reveal structural and functional details of DNA-binding proteins when subjected to X-ray diffraction. However, biomolecular crystals are notoriously unstable in solution conditions other than their native growth solution. To achieve greater application utility beyond structural biology, biomolecular crystals should be made robust against harsh conditions. To overcome this challenge, we optimized chemical DNA ligation within a co-crystal. Co-crystals from two distinct DNA-binding proteins underwent DNA ligation with the carbodiimide crosslinking agent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) under various optimization conditions: 5′ vs. 3′ terminal phosphate, EDC concentration, EDC incubation time, and repeated EDC dose. This crosslinking and DNA ligation route did not destroy crystal diffraction. In fact, the ligation of DNA across the DNA–DNA junctions was clearly revealed via X-ray diffraction structure determination. Furthermore, crystal macrostructure was fortified. Neither the loss of counterions in pure water, nor incubation in blood serum, nor incubation at low pH (2.0 or 4.5) led to apparent crystal degradation. These findings motivate the use of crosslinked biomolecular co-crystals for purposes beyond structural biology, including biomedical applications.

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Section: Discussionmentioning

confidence: 63%

Section: Introductionmentioning

confidence: 99%

Stabilizing DNA–Protein Co-Crystals via Intra-Crystal Chemical Ligation of the DNA

et al. 2021

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“…The crystalline matrix provides stable protein assemblies as a result of non‐covalent protein‐protein interactions [15] . The molecular arrangement in protein crystals has been used for enantioselective catalysis and for encapsulating organic and inorganic compounds with a wide range of sizes [15–23] . For this purpose, random cross‐linking induced by chemical reagents such as glutaraldehyde, has been used to prevent the decomposition of the crystals [15] …”

Section: Figurementioning

confidence: 99%

Design of an In‐Cell Protein Crystal for the Environmentally Responsive Construction of a Supramolecular Filament

et al. 2021

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Protein assemblies can be designed for development of nano–bio materials. This has been achieved by modulating protein–protein interactions. However, fabrication of highly ordered protein assemblies remains challenging. Protein crystals, which have highly ordered arrangements of protein molecules, provide useful source matrices for synthesizing artificial protein assemblies. Here, we describe construction of a supramolecular filament structure by engineering covalent and non‐covalent interactions in a protein crystal. Performing in‐cell crystallization of Trypanosoma brucei cysteine protease cathepsin B (TbCatB), we achieved a precise arrangement of protein molecules while suppressing random aggregation due to disulfide bonds. We succeeded in synthesizing bundled filament from the crystals by autoxidation of cysteinyl thiols after the isolation of the crystals from living cells.

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“…Our group has recently engineered metal coordination sites on the interior pore surfaces of CJ protein crystals (Figure 1b) to exert spatial and temporal control over macromolecular guest installation within the crystal scaffold (Figure 9c) (Huber, Hartje, McPherson, Kowalski, and Snow, 2017;Kowalski et al, 2016). We used stepwise guest loading and EDTA as a metal chelator to demonstrate secure immobilization and precise segregation.…”

Section: Engineering Crystal Surfaces and Pore Environmentsmentioning

confidence: 99%

“…Copyright 2016 American Chemical Society) (c) Confocal imaging of an interior plane within a highly porous CJ crystal, demonstrating spatially segregated macromolecular guests (mNeonGreen and mCherry) immobilized with Zn 2+ . (Reprinted with permission from Huber et al (2017). Copyright 2017 Royal Society of Chemistry) of shelf-stability (Shenoy et al, 2001) and decreased viscosity which could enable delivery via smaller needles (Basu et al, 2004).…”

Section: Pharmaceutical Formulationsmentioning

confidence: 99%

Protein crystal based materials for nanoscale applications in medicine and biotechnology

Hartje

Snow

2018

WIREs Nanomed Nanobiotechnol

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The porosity, order, biocompatibility, and chirality of protein crystals has motivated interest from diverse research domains including materials science, biotechnology, and medicine. Porous protein crystals have the unusual potential to organize guest molecules within highly ordered scaffolds, enabling applications ranging from biotemplating and catalysis to biosensing and drug delivery. Significant research has therefore been directed toward characterizing protein crystal materials in hopes of optimizing crystallization, scaffold stability, and application efficacy. In this overview article, we describe recent progress in the field of protein crystal materials with special attention given to applications in nanomedicine and nanobiotechnology.

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Programmed Assembly of Host–Guest Protein Crystals

Cited by 24 publications

References 13 publications

Stabilizing DNA–Protein Co-Crystals via Intra-Crystal Chemical Ligation of the DNA

Stabilizing DNA–Protein Co-Crystals via Intra-Crystal Chemical Ligation of the DNA

Design of an In‐Cell Protein Crystal for the Environmentally Responsive Construction of a Supramolecular Filament

Protein crystal based materials for nanoscale applications in medicine and biotechnology

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