title: High Resolution X-ray Diffraction Dataset for Bacillus licheniformis Gamma Glutamyl Transpeptidase-acivicin complex: SUMO-Tag Renders High Expression and Solubility.
abstract: Gamma glutamyl transpeptidase, (GGT) is a ubiquitous protein which plays a central role in glutathione metabolism and has myriad clinical implications. It has been shown to be a virulence factor for pathogenic bacteria, inhibition of which results in reduced colonization potential. However, existing inhibitors are effective but toxic and therefore search is on for novel inhibitors, which makes it imperative to understand the interactions of various inhibitors with the protein in substantial detail. High resolution structures of protein bound to different inhibitors can serve this purpose. Gamma glutamyl transpeptidase from Bacillus licheniformis is one of the model systems that have been used to understand the structure-function correlation of the protein. The structures of the native protein (PDB code 4OTT), of its complex with glutamate (PDB code 4OTU) and that of its precursor mimic (PDB code 4Y23) are available, although at moderate/low resolution. In the present study, we are reporting the preliminary analysis of, high resolution X-ray diffraction data collected for the co-crystals of B. licheniformis, Gamma glutamyl transpeptidase, with its inhibitor, Acivicin. Crystals belong to the orthorhombic space group P212121and diffract X-ray to 1.45 Å resolution. This is the highest resolution data reported for all GGT structures available till now. The use of SUMO fused expression system enhanced yield of the target protein in the soluble fraction, facilitating recovery of protein with high purity. The preliminary analysis of this data set shows clear density for the inhibitor, acivicin, in the protein active site.
authors.affiliation: Department of Biophysics, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India.
authors.forename: Shobha
authors.initials: S
authors.lastname: Kumari
authors.affiliation: National Institute of Immunology (NII), Aruna Asaf Ali Marg, New Delhi, 110067, India.
authors.forename: Ravi Kant
authors.initials: RK
authors.lastname: Pal
authors.affiliation: Department of Microbiology, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India.
authors.forename: Rani
authors.initials: R
authors.lastname: Gupta
authors.affiliation: Department of Biophysics, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India. manishagoel@south.du.ac.in.
authors.forename: Manisha
authors.initials: M
authors.lastname: Goel
mesh_terms: D000069976:Bacillus licheniformis / Q000201:enzymology*; D015870:Gene Expression; D007555:Isoxazoles / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D025842:SUMO-1 Protein / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction; D005723:gamma-Glutamyltransferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D007555:Isoxazoles; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; D005723:gamma-Glutamyltransferase; C006303:acivicin
keywords: Acivicin; Bacillus licheniformis; Gamma glutamyl transpeptidase; High resolution X-ray diffraction data; Inhibitor binding; SUMO-Tag expression system