Crystallization of a human Bence–Jones protein in microgravity using vapor diffusion in capillaries

Alvarado, U.R.; DeWitt, C.R.; Shultz, B.B.; Ramsland, Paul A.; Edmundson, Allen B.

doi:10.1016/s0022-0248(00)01011-3

Cited by 12 publications

(17 citation statements)

References 11 publications

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“…The evolution of our microgravity procedures into a relatively unsophisticated protocol culminated in the production of one crystal 12.8 mm 3 in volume, plus several hundred satellites, in 9 days without operator intervention on STS-95 (in October 1998). Morphologies of the microgravity-grown crystals were very similar to those in the ground-based forms (Alvarado et al, 2001;Bourne et al, 2002). As documented in the present work, however, it was necessary to confirm that the three-dimensional structures of the constituent Sea proteins were also identical in the two sets of crystals.…”

Section: Introductionsupporting

confidence: 59%

“…By methods previously described (Alvarado et al, 2001), the Bence-Jones dimer was prepared and purified from the urine of a patient (Sea) with multiple myeloma and amyloidosis. Samples used for crystallization were dialyzed into 0.05 M NaCl-0.05% sodium azide and concentrated by vacuum dialysis to 29-32 mg per ml.…”

Section: Experimental Preparation Purification and Crystallization Omentioning

confidence: 99%

“…The Sea protein does not readily form such large single crystals by batch methods. Under closely controlled conditions based on multiple transfers of preexisting crystals into fresh protein solutions (Shan et al, 1993), the Sea dimer has been found to form crystals with dimensions up to 1.3 Â 0.5 Â 0.3 mm in 13-30 days by partial dehydration of PEG 8000 solutions in sealed capillaries (Alvarado et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

“…Although highly successful, the intricate methods used for STS-80 needed to be simplified for comprehensive surveys of hundreds of samples. In the current procedure, the containment hardware is more convenient to assemble and the sample volumes can be varied to accelerate the loading process and to improve the reproducibility of the results (Alvarado et al, 2001). The evolution of our microgravity procedures into a relatively unsophisticated protocol culminated in the production of one crystal 12.8 mm 3 in volume, plus several hundred satellites, in 9 days without operator intervention on STS-95 (in October 1998).…”

Section: Introductionmentioning

confidence: 99%

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Comparison of the three‐dimensional structures of a human Bence‐Jones dimer crystallized on Earth and aboard US Space Shuttle Mission STS‐95

Terzyan

Ramsland

Bourne

et al. 2003

J of Molecular Recognition

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Crystals of a human (Sea) Bence-Jones dimer were produced in a capillary by vapor diffusion under microgravity conditions in the 9 day US Space Shuttle Mission STS-95. In comparison to ground-based experiments, nucleation was facile and spontaneous in space. Appearance of a very large (8 x 1.6 x 1.0 mm) crystal in a short time period is a strong endorsement for the use of microgravity to produce crystals sufficiently large for neutron diffraction studies. The Sea dimer crystallized in the orthorhombic space group P2(1)2(1)2(1), with a = 48.9 A, b = 85.2 A, and c = 114.0 A. The crystals grown in microgravity exhibited significantly lower mosaicities than those of ground-based crystals and the X-ray diffraction data had a lower overall B factor. Three-dimensional structures determined by X-ray analysis at two temperatures (100 and 293 K) were indistinguishable from those obtained from ground-based crystals. However, both the crystallographic R factor and the free R factor were slightly lower in the models derived from crystals produced in microgravity. The major difference between the two crystal growth systems is a lack of convection and sedimentation in a microgravity environment. This environment resulted in the growth of much larger, higher-quality crystals of the Sea Bence-Jones protein. Structurally, heretofore unrecognized grooves on the external surfaces of the Sea and other immunoglobulin-derived fragments are regular features and may offer supplementary binding regions for super antigens and other elongated ligands in the bloodstream and perivascular tissues.

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Section: Introductionsupporting

confidence: 59%

Section: Experimental Preparation Purification and Crystallization Omentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparison of the three‐dimensional structures of a human Bence‐Jones dimer crystallized on Earth and aboard US Space Shuttle Mission STS‐95

Terzyan

Ramsland

Bourne

et al. 2003

J of Molecular Recognition

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“…Particularly, amyloidogenic light chains are commonly found to be in equilibrium between the dimer and the monomeric form in patients' urine [16]. After the production and purification step, the protein has been characterized in terms of monomer-dimer equilibrium at physiological and low pH.…”

Section: Resultsmentioning

confidence: 99%

In Vitro Aggregation Behavior of a Non-Amyloidogenic λ Light Chain Dimer Deriving from U266 Multiple Myeloma Cells

et al. 2012

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Excessive production of monoclonal light chains due to multiple myeloma can induce aggregation-related disorders, such as light chain amyloidosis (AL) and light chain deposition diseases (LCDD). In this work, we produce a non-amyloidogenic IgE λ light chain dimer from human mammalian cells U266, which originated from a patient suffering from multiple myeloma, and we investigate the effect of several physicochemical parameters on the in vitro stability of this protein. The dimer is stable in physiological conditions and aggregation is observed only when strong denaturating conditions are applied (acidic pH with salt at large concentration or heating at melting temperature Tm at pH 7.4). The produced aggregates are spherical, amorphous oligomers. Despite the larger β-sheet content of such oligomers with respect to the native state, they do not bind Congo Red or ThT. The impossibility to obtain fibrils from the light chain dimer suggests that the occurrence of amyloidosis in patients requires the presence of the light chain fragment in the monomer form, while dimer can form only amorphous oligomers or amorphous deposits. No aggregation is observed after denaturant addition at pH 7.4 or at pH 2.0 with low salt concentration, indicating that not a generic unfolding but specific conformational changes are necessary to trigger aggregation. A specific anion effect in increasing the aggregation rate at pH 2.0 is observed according to the following order: SO4 −≫Cl−>H2PO4 −, confirming the peculiar role of sulfate in promoting protein aggregation. It is found that, at least for the investigated case, the mechanism of the sulfate effect is related to protein secondary structure changes induced by anion binding.

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Drug Design

Betzel¹,

Martirosyan²

2017

Biotechnology in Space

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Crystallization of a human Bence–Jones protein in microgravity using vapor diffusion in capillaries

Cited by 12 publications

References 11 publications

Comparison of the three‐dimensional structures of a human Bence‐Jones dimer crystallized on Earth and aboard US Space Shuttle Mission STS‐95

Comparison of the three‐dimensional structures of a human Bence‐Jones dimer crystallized on Earth and aboard US Space Shuttle Mission STS‐95

In Vitro Aggregation Behavior of a Non-Amyloidogenic λ Light Chain Dimer Deriving from U266 Multiple Myeloma Cells

Drug Design

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