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Heterogeneous nucleation helps the search for initial crystallization conditions of ?-glutamyl transpeptidase from Bacillus licheniformis.

ABSTRACT: Here, the crystallization and preliminary X-ray diffraction studies of Bacillus licheniformis ?-glutamyl transpeptidase (BlGT) are reported. The serendipitous finding of heterogeneous nucleants in the initial experiments provided the first crystallization conditions for the protein. Crystals were grown by hanging-drop vapour diffusion using a precipitant solution consisting of 20%(w/v) PEG 3350, 0.2 M magnesium chloride hexahydrate, 0.1 M Tris-HCl pH 8.2. The protein crystallized in the orthorhombic space group P2(1)2(1)2(1), with one heterodimer per asymmetric unit and unit-cell parameters a = 60.90, b = 61.97, c = 148.24 Å. The BlGT crystals diffracted to 2.95 Å resolution.

SUBMITTER: Lin LL

PROVIDER: S-EPMC3668591 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Heterogeneous nucleation helps the search for initial crystallization conditions of γ-glutamyl transpeptidase from Bacillus licheniformis.

Lin Long Liu LL Merlino Antonello A

Acta crystallographica. Section F, Structural biology and crystallization communications 20130525 Pt 6

Here, the crystallization and preliminary X-ray diffraction studies of Bacillus licheniformis γ-glutamyl transpeptidase (BlGT) are reported. The serendipitous finding of heterogeneous nucleants in the initial experiments provided the first crystallization conditions for the protein. Crystals were grown by hanging-drop vapour diffusion using a precipitant solution consisting of 20%(w/v) PEG 3350, 0.2 M magnesium chloride hexahydrate, 0.1 M Tris-HCl pH 8.2. The protein crystallized in the orthorho ...[more]

PMID: 23722850

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Covalent Immobilization of Bacillus licheniformis ?-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles.

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Statistical optimization of culture conditions of mesophillic gamma-glutamyl transpeptidase from <i>Bacillus altitudinis</i> IHB B1644.

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2023-07-05 | GSE217834 | GEO

The clinical implication of gamma-glutamyl transpeptidase in COVID-19.

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Human lung expresses unique gamma-glutamyl transpeptidase transcripts.

Project description:gamma-Glutamyl transpeptidase (EC 2.3.2.2, gamma GT) is a membrane-bound ectoenzyme that plays an important role in the metabolism of glutathione. It is composed of two subunits, both of which are encoded by a common mRNA. We examined the expression of gamma GT in human lung tissue by Northern blot analysis and screening a cDNA library made from human lung poly(A)+ RNA. Our results show that there are two gamma GT mRNA populations in human lung tissue. We define these as group I (2.4 kb) and group II (approximately 1.2 kb) transcripts. In the present communication, we characterize the unique lung transcript. Sequence analysis of representative clones shows that group I transcripts are virtually identical to those previously isolated from liver and placenta but possess a unique 5' untranslated region. In marked contrast, group II transcripts appear to be human-lung-specific. Group II transcripts appear on Northern blots probed with full-length or 3'-biased gamma GT cDNA. Sequence analysis of group II clones shows them to be homologous with group I clones in the region that encodes the reading frame for the light chain; however, they possess a series of unique 5' untranslated regions, which suggests that they arise from lung-specific message processing. Additionally, approximately 50% of the isolated group II clones contain 34 nt substitutions compared with the "wild-type" gamma GT transcripts. These data indicate that human lung expresses unique gamma GT transcripts of unknown function as well as the classical form. The abundant group II transcripts may encode part of a heterodimer related to gamma GT or represent processed lung-specific pseudogenes.

| S-EPMC47161 | biostudies-other

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