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Effect of Amino Acid Substitution in the Penaeus monodon LGBP and Specificity Through Mutational Analysis

  • Jeyachandran SivakamavalliEmail author
  • Chandrabose Selvaraj
  • Sanjeev Kumar Singh
  • Kiyun Park
  • Ihn-Sil Kwak
  • Baskaralingam VaseeharanEmail author
Article
  • 35 Downloads

Abstract

Lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) is a pattern recognition protein (PRP) purified from the Penaeus monodon by Blue-Sepharose, Phenyl-Sepharose followed by Sephadex G-100 chromatography. P. monodon LGBP consist of 36 and 48 kDa subunits on 10% SDS-PAGE under reducing and non-reducing conditions respectively. Purified P. monodon LGBP agglutinates the fungal pathogen Candida glabrata, due to the presence of β-glucan (βG) on its surface. This agglutination was cross checked with the in silico docking analysis of LGBP-βG and LGBP-laminarin (isomeric form βG) interaction. As part of a strategy, to determine the precise role of P. monodon LGBP (Pm-LGBP) in pattern recognition mechanism mutations were introduced by in silico approach. In crustacean LGBP, RGD motif (Arg, Gly, Asp) plays the vital role in the cell adhesion and pattern recognition mechanism. Role of Asp in RGD motif was determined through amino acid substitution, introduction of a specific mutation D134K into a central area of the sugar-binding (βG) site resulted in complete loss of pathogen recognition and binding of Pm-LGBP to βG. These results demonstrate that, the RGD motif of Asp134 is essential for sugar binding in P. monodon. To our knowledge, P. monodon D134K is the first mutant shrimp LGBP which is unable to bind with the sugar residues. This mutant could be useful in the discovery of actual function of Pm-LGBP in the recognition of homologous symbionts.

Graphic Abstract

Docking analysis of LGBP binding towards β-glucan (i) LGBP without mutationv (ii) LGBP with mutation.

Keywords

Docking Homology modelling LGBP Molecular simulation Mutation RMSD SDS-PAGE 

Notes

Acknowledgements

The study was supported by the National Research Foundation of Korea, which is funded by the Korean Government [NRF-2018-R1A6A1A-03024314]. The authors Chandrabose Selvaraj and Sanjeev Kumar Singh thankfully acknowledge financial support of RUSA-Phase 2.0 grant sanctioned vide Letter No: F.24-51/2014-U, Policy (TNmulti-Gen), Dept. of Edn, Govt. of India, Dt.09.10.2018.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10989_2019_9960_MOESM1_ESM.doc (184 kb)
Supplementary file1 (DOC 184 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Jeyachandran Sivakamavalli
    • 1
    • 2
    Email author
  • Chandrabose Selvaraj
    • 3
  • Sanjeev Kumar Singh
    • 3
  • Kiyun Park
    • 1
  • Ihn-Sil Kwak
    • 1
    • 4
  • Baskaralingam Vaseeharan
    • 2
    Email author
  1. 1.Department of Fisheries Research InstituteChonnam National UniversityYeosuSouth Korea
  2. 2.Crustacean Molecular Biology and Genomics Lab, Department of Animal Health and ManagementAlagappa UniversityKaraikudiIndia
  3. 3.Computer Aided Drug Designing and Molecular Modeling Lab, Department of BioinformaticsAlagappa UniversityKaraikudiIndia
  4. 4.Faculty of Marine TechnologyChonnam National UniversityChonnamRepublic of Korea

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