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Highly efficient site-specific protein modification using tyrosinase from Streptomyces avermitilis: Structural

Hwaseok Hong1, Uk-Jae Lee2, Seul Hoo Lee1

  • 1School of Life Sciences, BK21 FOUR KNU Creative BioResearch Group, KNU Institute of Microbiology, Kyungpook National University, Daegu 41566, Republic of Korea.

International Journal of Biological Macromolecules
|November 23, 2023
PubMed
Summary

A novel tyrosinase, SaTYR, enables site-specific protein modification by reacting with surface tyrosine residues more effectively than previously known enzymes. This expands protein bioconjugation possibilities for various applications.

Keywords:
CrosslinkingProtein bioconjugationTyrosinase

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Area of Science:

  • Biochemistry
  • Protein Engineering
  • Enzymology

Background:

  • Tyrosinase-mediated protein conjugation is a mild, site-specific modification method.
  • Existing tyrosinases are limited to highly exposed tyrosine residues, restricting modification sites.

Purpose of the Study:

  • To identify and characterize a novel tyrosinase with enhanced activity on surface tyrosine residues.
  • To expand the utility of tyrosinase-mediated protein modification.

Main Methods:

  • Enzyme discovery and characterization of tyrosinase from Streptomyces avermitilis (SaTYR).
  • Crystal structure determination of SaTYR.
  • Demonstration of SaTYR-mediated fluorescence dye tagging and PEGylation on surface tyrosine residues.

Main Results:

  • SaTYR exhibits significantly higher activity on surface tyrosine residues compared to other tyrosinases.
  • Crystal structure reveals a flat, shallow substrate-binding pocket suitable for protein substrates.
  • Achieved ~95.2% conjugation yield in 1 hour for previously unreacted surface tyrosine residues.
  • Provided a structural rationale for selecting optimal tyrosine target sites.

Conclusions:

  • SaTYR overcomes limitations of previous tyrosinases, enabling modification of a wider range of surface tyrosine residues.
  • SaTYR significantly broadens the scope of tyrosinase-mediated protein modification.
  • Site-specific tyrosine conjugation using SaTYR is a promising strategy for protein bioconjugation.