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High Resolution X-ray Diffraction Dataset for Bacillus licheniformis Gamma Glutamyl Transpeptidase-acivicin complex: SUMO-Tag Renders High Expression and Solubility

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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 P212121 and 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.

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Abbreviations

GGT:

Gamma glutamyl transpeptidase

BlGGT:

Bacillus licheniformis gamma glutamyl transpeptidase

IPTG:

Isopropyl-thio-β-d-galactoside

Ni–NTA:

Nickel-nitrilotriacetic acid

PEG:

Polyethylene glycol

SDS–PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

FT:

Flow through

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Acknowledgements

This work was supported by CSIR (Council of Scientific and Industrial Research), Government of India through Grant No. 38 (1312)/11/EMR-II to the PI (Dr. Manisha Goel). Shobha Kumari also acknowledges the CSIR-JRF fellowship (09/045(1094)/2011-EMR-1). The authors gratefully acknowledge National Institute of Immunology, New Delhi for providing X-ray diffraction data collection facility. The authors also acknowledge the CIF (Central Instrumentation Facility) at University of Delhi South Campus for MALDI studies.

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Authors and Affiliations

  1. Department of Biophysics, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Shobha Kumari & Manisha Goel

  2. National Institute of Immunology (NII), Aruna Asaf Ali Marg, New Delhi, 110067, India

    Ravi Kant Pal

  3. Department of Microbiology, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Rani Gupta

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  1. Shobha Kumari
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Correspondence to Manisha Goel.

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Kumari, S., Pal, R.K., Gupta, R. et al. High Resolution X-ray Diffraction Dataset for Bacillus licheniformis Gamma Glutamyl Transpeptidase-acivicin complex: SUMO-Tag Renders High Expression and Solubility. Protein J 36, 7–16 (2017). https://doi.org/10.1007/s10930-017-9693-2

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