Protein crystallization: Eluding the bottleneck of X-ray crystallography

Holcomb, Joshua; Spellmon, Nicholas; Zhang, Yingxue; Doughan, Maysaa; Li, Chunying; Yang, Zhe

doi:10.3934/biophy.2017.4.557

Cited by 38 publications

(28 citation statements)

References 103 publications

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“…It can take researchers many years to find a way around these challenges in order to crystallize certain specimen but there has been progress made in the field. It has been found that the addition of additives such as small molecules, detergents or metal ions facilitate or enhance crystal formation or growth [10]. In 2006, Tomcova etal.…”

Section: X-ray Crystallographymentioning

confidence: 99%

Seeing is believing: structures and functions of biological molecules

Boodhun

2018

BioTechniques

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Section: X-ray Crystallographymentioning

confidence: 99%

Seeing is believing: structures and functions of biological molecules

Boodhun

2018

BioTechniques

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“…The goal of macromolecular crystallography is to reveal the macromolecule's three-dimensional (3D) structure at the atom's level by using X-ray diffraction on crystals obtained from a variety of experimental setups 16 . Crystallization in space begun~40 years ago with the idea that in microgravity the crystals will grow with enhanced properties that would eventually improve the quality of the derived X-ray diffraction data 17,18 .…”

Section: Introductionmentioning

confidence: 99%

Protein structural changes on a CubeSat under rocket acceleration profile

et al. 2020

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Catalyzing life-sustaining reactions, proteins are composed by 20 different amino acids that fold into a compact yet flexible threedimensional architecture, which dictates what their function(s) might be. Determining the spatial arrangement of the atoms, the protein's 3D structure, enables key advances in fundamental and applied research. Protein crystallization is a powerful technique to achieve this. Unlike Earth's crystallization experiments, biomolecular crystallization in space in the absence of gravitational force is actively sought to improve crystal growth techniques. However, the effects of changing gravitational vectors on a protein solution reaching supersaturation remain largely unknown. Here, we have developed a low-cost crystallization cell within a CubeSat payload module to exploit the unique experimental conditions set aboard a sounding rocket. We designed a biaxial gimbal to house the crystallization experiments and take measurements on the protein solution in-flight with a spectrophotometry system. After flight, we used X-ray diffraction analysis to determine that flown protein has a structural rearrangement marked by loss of the protein's water and sodium in a manner that differs from crystals grown on the ground. We finally show that our gimbal payload module design is a portable experimental setup to take laboratory research investigations into exploratory space flights.

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“…A majority of the established protein structures have been determined by X-ray crystallography, because it is a well-established methodology for solving protein structures with a large span of molecular weights [2,6,7]. To obtain crystal structures, it is required to experimentally obtain high-quality protein crystals for the X-ray diffraction experiments, which is proven to be the bottleneck for many proteins [8,9], especially for those large proteins or protein complexes that are over 100 kDa [10][11][12]. It is estimated that of all of the proteins used for crystallization trials, only one-third can be crystallized, of which about a half can further produce usable-quality X-ray diffraction data through devising and diversifying the proper optimization of crystallization conditions (http://targetdb.pdb.org) [13,14].…”

Section: Introductionmentioning

confidence: 99%

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

Zhang

Wang

Jia

2020

IJMS

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Although prevalent in the determination of protein structures; crystallography always has the bottleneck of obtaining high-quality protein crystals for characterizing a wide range of proteins; especially large protein complexes. Stable fragments or domains of proteins are more readily to crystallize; which prompts the use of in situ proteolysis to remove flexible or unstable structures for improving crystallization and crystal quality. In this work; we investigated the effects of in situ proteolysis by chymotrypsin on the crystallization of the XcpVWX complex from the Type II secretion system of Pseudomonas aeruginosa. Different proteolysis conditions were found to result in two distinct lattices in the same crystallization solution. With a shorter chymotrypsin digestion at a lower concentration; the crystals exhibited a P3 hexagonal lattice that accommodates three complex molecules in one asymmetric unit. By contrast; a longer digestion with chymotrypsin of a 10-fold higher concentration facilitated the formation of a compact P212121 orthorhombic lattice with only one complex molecule in each asymmetric unit. The molecules in the hexagonal lattice have shown high atomic displacement parameter values compared with the ones in the orthorhombic lattice. Taken together; our results clearly demonstrate that different proteolysis conditions can result in the generation of distinct lattices in the same crystallization solution; which can be exploited in order to obtain different crystal forms of a better quality

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Protein crystallization: Eluding the bottleneck of X-ray crystallography

Cited by 38 publications

References 103 publications

Seeing is believing: structures and functions of biological molecules

Seeing is believing: structures and functions of biological molecules

Protein structural changes on a CubeSat under rocket acceleration profile

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

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