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Updated: Jul 16, 2026
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OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
Published on: February 5, 2020
Sortase A as a novel molecular "stapler" for sequence-specific protein conjugation.
Ranganath Parthasarathy1, Shyamsundar Subramanian, Eric T Boder
1Departments of Chemical and Biomolecular Engineering and Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Bioconjugate Chemistry
|February 17, 2007
Summary
Sortase A enzyme enables precise protein attachment to surfaces and materials. This bacterial enzyme facilitates protein modification and immobilization for various applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Materials Science
Background:
- Sortase enzymes are bacterial transpeptidases crucial for cell wall anchoring.
- Sortase A from Staphylococcus aureus is a well-characterized enzyme in this family.
Purpose of the Study:
- To explore the utility of recombinant Sortase A for protein immobilization and modification.
- To investigate Sortase A's ability to ligate proteins to synthetic materials and generate protein constructs.
Main Methods:
- Utilized recombinant Staphylococcus aureus Sortase A enzyme.
- Attached a model protein substrate (green fluorescent protein, eGFP) to functionalized polystyrene beads.
- Investigated ligation to amino-terminated poly(ethylene glycol) and generation of protein oligomers/cyclized monomers.
Main Results:
- Successfully attached eGFP to polystyrene beads modified with alkylamine or Gly-Gly-Gly.
- Demonstrated Sortase A-mediated ligation of eGFP to poly(ethylene glycol).
- Showed the enzyme's capability to create protein oligomers and cyclized monomers.
- Identified alkylamine as a viable alternative to the natural Gly3 substrate.
Conclusions:
- Sortase A is a versatile tool for sequence-specific protein immobilization and modification.
- The enzyme's mild reaction conditions and specific recognition tags are advantageous.
- Alkylamine substitution suggests potential for Sortase A in materials science applications.
- Sortase A expands the toolkit for protein engineering and surface functionalization.
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