Complexes of cysteine proteases with chagasin

Bujacz, G.; Redzynia, I.; Bujacz, A.; Jaskólski, M.; Abrahamson, Magnus; Ljunggren, Anna

doi:10.1107/s0108767309097086

Cited by 2 publications

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“…The ESA crystals were grown from ammonium sulfate and required the use of 30% glycerol as a cryoprotectant. Crystals obtained in the presence of 70% Tacsimate were mounted without additional cryoprotection, confirming the fact that carboxylate salts present in the crystallization drop can successfully act as cryoprotective agents during diffraction experiments (Bujacz et al, 2010). These new conditions were not only highly cryoprotective, but also improved the initial resolution from 2.32 to 2.04 Å .…”

Section: X-ray Diffraction Data Collectionmentioning

confidence: 54%

Structures of bovine, equine and leporine serum albumin

Bujacz

2012

Acta Crystallogr D Biol Crystallogr

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Serum albumin first appeared in early vertebrates and is present in the plasma of all mammals. Its canonical structure supported by a conserved set of disulfide bridges is maintained in all mammalian serum albumins and any changes in sequence are highly correlated with evolution of the species. Previous structural investigations of mammalian serum albumins have only concentrated on human serum albumin (HSA), most likely as a consequence of crystallization and diffraction difficulties. Here, the crystal structures of serum albumins isolated from bovine, equine and leporine blood plasma are reported. The structure of bovine serum albumin (BSA) was determined at 2.47 Å resolution, two crystal structures of equine serum albumin (ESA) were determined at resolutions of 2.32 and 2.04 Å, and that of leporine serum albumin (LSA) was determined at 2.27 Å resolution. These structures were compared in detail with the structure of HSA. The ligand-binding pockets in BSA, ESA and LSA revealed different amino-acid compositions and conformations in comparison to HSA in some cases; however, much more significant differences were observed on the surface of the molecules. BSA, which is one of the most extensively utilized proteins in laboratory practice and is used as an HSA substitute in many experiments, exhibits only 75.8% identity compared with HSA. The higher resolution crystal structure of ESA highlights the binding properties of this protein because it includes several bound compounds from the crystallization solution that provide additional structural information about potential ligand-binding pockets.

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Section: X-ray Diffraction Data Collectionmentioning

confidence: 54%

Structures of bovine, equine and leporine serum albumin

Bujacz

2012

Acta Crystallogr D Biol Crystallogr

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690

554

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“…Sitting drops of PilA1Á1-34 were dispensed in 1:1 and 2:1 protein:crystallization solution ratios in MRC crystallization plates with a Mosquito dispensing robot (TTP Labtech) using several commercial crystallization screens. Crystals measuring 150 Â 150 Â 100 mm were obtained after four days of incubation at 20 C in 1.6 M sodium citrate pH 6.5, a condition that provides cryoprotection (Bujacz et al, 2010). Crystals were harvested in nylon loops and directly flash-cooled in liquid nitrogen for subsequent synchrotron data collection (Garman & Owen, 2006).…”

Section: Crystallizationmentioning

confidence: 99%

A practical overview of molecular replacement: Clostridioides difficile PilA1, a difficult case study

Crawshaw

Baslé

Salgado

2020

Acta Cryst Sect D Struct Biol

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Many biologists are now routinely seeking to determine the three‐dimensional structures of their proteins of choice, illustrating the importance of this knowledge, but also of the simplification and streamlining of structure‐determination processes. Despite the fact that most software packages offer simple pipelines, for the non‐expert navigating the outputs and understanding the key aspects can be daunting. Here, the structure determination of the type IV pili (TFP) protein PilA1 from Clostridioides difficile is used to illustrate the different steps involved, the key decision criteria and important considerations when using the most common pipelines and software. Molecular‐replacement pipelines within CCP4i2 are presented to illustrate the more commonly used processes. Previous knowledge of the biology and structure of TFP pilins, particularly the presence of a long, N‐terminal α‐helix required for pilus formation, allowed informed decisions to be made during the structure‐determination strategy. The PilA1 structure was finally successfully determined using ARCIMBOLDO and the ab initio MR strategy used is described.

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Complexes of cysteine proteases with chagasin

Cited by 2 publications

References 0 publications

Structures of bovine, equine and leporine serum albumin

Structures of bovine, equine and leporine serum albumin

A practical overview of molecular replacement: Clostridioides difficile PilA1, a difficult case study

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