1. Academic Validation
  2. Molecular cloning and biophysical characterization of CXCL3 chemokine

Molecular cloning and biophysical characterization of CXCL3 chemokine

  • Int J Biol Macromol. 2018 Feb;107(Pt A):575-584. doi: 10.1016/j.ijbiomac.2017.09.032.
Khushboo Gulati 1 Krishnakant Gangele 1 Nipanshu Agarwal 1 Minal Jamsandekar 1 Dinesh Kumar 2 Krishna Mohan Poluri 3
Affiliations

Affiliations

  • 1 Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, 247667 Uttarakhand, India.
  • 2 Centre for Biomedical Research, SGPGIMS Campus, Lucknow 226014, Uttar Pradesh, India.
  • 3 Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, 247667 Uttarakhand, India; Centre for Nanotechnology, Indian Institute of Technology Roorkee, Roorkee, 247667 Uttarakhand, India. Electronic address: krishfbt@iitr.ac.in.
Abstract

CXCL3 is a neutrophil activating chemokine that belongs to GRO subfamily of CXC Chemokines. GRO chemokine family comprises of three chemokines GRO α (CXCL1), GROβ (CXCL2), and GRO γ (CXCL3), which arose as a result of gene duplication events during the course of chemokine evolution. Although primary sequences of GRO chemokines are highly similar, they performs several protein specific functions in addition to their common property of neutrophil trafficking. However, the molecular basis for their differential functions has not well understood. Although structural details are available for CXCL1 and CXCL2, no such information regarding CXCL3 is available till date. In the present study, we have successfully cloned, expressed, and purified the recombinant CXCL3. Around 15mg/L of pure recombinant CXCL3 protein was obtained. Further, we investigated its functional divergence and biophysical characteristics such as oligomerization, thermal stability and heparin binding etc., and compared all these features with its closest paralog CXCL2. Our studies revealed that, although overall structural and oligomerization features of CXCL3 and CXCL2 are similar, prominent differences were observed in their surface characteristics, thus implicating for a functional divergence.

Keywords

CXCL3; Functional divergence; GRO chemokines; Heparin binding; Homodimer; Molecular evolution.

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