Functions, structures and Triton X-100 effect for the catalytic subunits of heterodimeric phospholipases A2 from Vipera nikolskii venom.

Phospholipases A(2) (PLA(2)s) from snake venoms have diverse pharmacological functions including neurotoxicity, and more studies are necessary to understand relevant mechanisms. Here we report the different crystal structures for two enzymatically active basic subunits (HDP-1P and HDP-2P) of heterodimeric neurotoxic PLA(2)s isolated from Vipera nikolskii venom. Structural comparisons with similar PLA(2)s clearly show some flexible regions which might be important for the catalytic function and neurotoxicity.

Octameric structure of the human bifunctional enzyme PAICS in purine biosynthesis.

Phosphoribosylaminoimidazole carboxylase/phosphoribosylaminoimidazole succinocarboxamide synthetase (PAICS) is an important bifunctional enzyme in de novo purine biosynthesis in vertebrate with both 5-aminoimidazole ribonucleotide carboxylase (AIRc) and 4-(N-succinylcarboxamide)-5-aminoimidazole ribonucleotide synthetase (SAICARs) activities. It becomes an attractive target for rational anticancer drug design, since rapidly dividing cancer cells rely heavily on the purine de novo pathway for synthesis of adenine and guanine, whereas normal cells favor the salvage pathway. Here, we report the crystal structure of human PAICS, the first in the entire PAICS family, at 2.

Crystallization and preliminary crystallographic studies of human septin 1 with site-directed mutations.

Septin 1 is a member of an evolutionarily conserved family of GTP-binding and filament-forming proteins named septins, which function in diverse processes including cytokinasis, vesicle trafficking, apoptosis, remodelling of the cytoskeleton, infection, neurodegeneration and neoplasia. Human septin 1 has been expressed and purified, but suffers from severe aggregation. Studies have shown that septin 1 with site-directed mutations of five serine residues (Ser19, Ser206, Ser307, Ser312 and Ser315) has a much lower degree of aggregation and better structural homogeneity and that the mutations cause only slight perturbations in the secondary structure of septin 1.

Crystallization and preliminary X-ray crystallographic studies of human cyclophilin J.

Human cyclophilin J, a new member of the cyclophilin family, has been expressed and crystallized. Diffraction data have been collected to 2.0 A resolution and preliminary crystallographic studies have been completed.

Isolation and preliminary crystallographic studies of two new phospholipases A2 from Vipera nikolskii venom.

Snake-venom phospholipases A2 (PLA2s) represent a good model for studies of structure-function relationships, mainly because of their small size and diverse pharmacological and toxicological activities. To obtain new members of the abundant PLA2 family, the venom of the viper Vipera nikolskii was fractionated for the first time and two new proteins, VN5-3 and VN4-3, were isolated. Both proteins show phospholipase A2 activity and may possess neurotoxic activity.

An attempt to increase the efficiency of protein crystal screening: a simplified screen and experiments.

Macromolecular crystallization remains a bottleneck in structure determination by X-ray diffraction. Based on the data reflecting success rates of crystallization conditions in different screens and the information derived from the BMCD and other related studies, a simplified screen has been designed to increase the success rate of traditional screening and to save samples, time and cost. The screen has been tested with six protein samples which had been crystallized before and its comparison with Crystal Screen (Hampton Research) was also performed with lysozyme crystallization.

2.6 A resolution crystal structure of the bacterioferritin from Azotobacter vinelandii.

The crystal structure of the bacterioferritin from Azotobacter vinelandii has been determined at 2.6 A resolution. Both the low occupancy of one iron ion in the dinuclear iron center and the deviation of its adjacent residue His130 from the center suggest migration of the iron ion from the dinuclear iron site to the inner nucleation site.

Purification and refolding of human alpha5-subunit (PSMA5) of the 20S proteasome, expressed as inclusion bodies in Escherichia coli.

The 20S proteasome is the central enzyme of nonlysosomal protein degradation in both the cytosol and nucleus. It is composed of 28 protein subunits which are arranged into four staggered heptameric rings. The outer rings consist of alpha-subunits which are responsible for binding of proteasome activators, inhibitors, and regulators.

The space experiment of protein crystallization aboard the Chinese spacecraft SZ-3.

Using new flight hardware, a Chinese mission of space protein crystallization has been performed aboard the Chinese spacecraft SZ-3. Preliminary analyses of the experimental results have shown that a few proteins produced better crystals in space. At least, the crystals of cytochrome b5 mutant could diffract X-ray beyond the highest resolution reported so far for the same kind of crystals.