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Oriented Immobilization on Gold Nanoparticles of a Recombinant Therapeutic Zymogen

  • Elina Dosadina
  • Celetia Agyeiwaa
  • William Ferreira
  • Simon Cutting
  • Abdullah Jibawi
  • Enrico Ferrari
  • Mikhail SolovievEmail author
Protocol
  • 156 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2118)

Abstract

Direct immobilization of functional proteins on gold nanoparticles (AuNPs) affects their structure and function. Changes may vary widely and range from strong inhibition to the enhancement of protein function. More often though the outcome of direct protein immobilization results in protein misfolding and the loss of protein activity. Additional complications arise when the protein being immobilized is a zymogen which requires and relies on additional protein–protein interactions to exert its function. Here we describe molecular design of a glutathione-S-transferase-Staphylokinase fusion protein (GST-SAK) and its conjugation to AuNPs. The multivalent AuNP-(GST-SAK)n complexes generated show plasminogen activation activity in vitro. The methods described are transferable and could be adapted for conjugation and functional analysis of other plasminogen activators, thrombolytic preparations or other functional enzymes.

Key words

Gold nanoparticles Plasminogen activator Therapeutic protein Protein function Protein stability Experimental design Protein immobilization Staphylokinase Oriented immobilization 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Elina Dosadina
    • 1
  • Celetia Agyeiwaa
    • 1
  • William Ferreira
    • 1
  • Simon Cutting
    • 1
  • Abdullah Jibawi
    • 2
  • Enrico Ferrari
    • 3
  • Mikhail Soloviev
    • 4
    Email author
  1. 1.Centre for Biomedical Sciences, School of Biological SciencesRoyal Holloway University of LondonEghamUK
  2. 2.Ashford and St. Peter’s Hospitals NHS Foundation TrustSurreyUK
  3. 3.College of Science, School of Life SciencesUniversity of Lincoln, Brayford PoolLincolnUK
  4. 4.Centre for Biomedical Sciences, Department of Biological SciencesRoyal Holloway University of LondonEghamUK

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