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Recent progress in enzymatic protein labelling techniques and their applications.

Yi Zhang1, Keun-Young Park, Kiall F Suazo

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Enzymatic protein labeling offers a highly efficient and robust method for creating diverse protein conjugates. This review highlights advancements in enzymatic techniques over five years for applications in therapeutics, imaging, and biosensing.

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Area of Science:

  • Bioconjugation Chemistry
  • Protein Engineering
  • Biotechnology

Background:

  • Protein conjugates are crucial for applications in diagnostics, therapeutics, and research.
  • Traditional chemical modification methods often result in heterogeneous products, complicating characterization and potentially affecting protein function.
  • Site-specific modification is essential for controlled protein engineering and functionalization.

Purpose of the Study:

  • To review recent advancements (last five years) in site-specific enzymatic protein labeling.
  • To highlight the efficiency and robustness of enzymatic methods compared to traditional chemical approaches.
  • To showcase the diverse applications of enzymatically labeled proteins.

Main Methods:

  • Review of literature focusing on enzymatic protein labeling techniques.
  • Analysis of developments in the application of ten specific enzymes: sortase A, subtiligase, microbial transglutaminase, farnesyltransferase, N-myristoyltransferase, phosphopantetheinyl transferases, tubulin tyrosin ligase, lipoic acid ligase, biotin ligase, and formylglycine generating enzyme.
  • Summarization of progress in creating protein conjugates for various applications.

Main Results:

  • Enzymatic protein labeling provides efficient and robust site-specific modification under mild conditions.
  • Significant progress has been made in applying enzymatic methods for creating clinically relevant conjugates, imaging agents, and probes.
  • Enzymatic approaches overcome limitations of traditional chemical modifications and unnatural amino acid incorporation.

Conclusions:

  • Enzymatic protein labeling is a powerful strategy for generating homogeneous and functional protein conjugates.
  • Recent developments have expanded the utility of enzymatic methods across therapeutics, diagnostics, and biosensing.
  • This review underscores the growing importance and versatility of enzymatic protein modification in biotechnology.