Protein-Protein Fusion Catalyzed by Sortase A

Chimeric proteins boast widespread use in areas ranging from cell biology to drug delivery. Post-translational protein fusion using the bacterial transpeptidase sortase A provides an attractive alternative when traditional gene fusion fails. We describe use of this enzyme for in vitro protein ligation and report the successful fusion of 10 pairs of protein domains with preserved functionality — demonstrating the robust and facile nature of this reaction.

[1]  S. Mazmanian,et al.  Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus , 2002, The Journal of Biological Chemistry.

[2]  S. Mazmanian,et al.  Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus , 2002, The Journal of Biological Chemistry.

[3]  J. Tom,et al.  A designed peptide ligase for total synthesis of ribonuclease A with unnatural catalytic residues. , 1994, Science.

[4]  K. Wittrup,et al.  Stability and CDR composition biases enrich binder functionality landscapes. , 2010, Journal of molecular biology.

[5]  S. Mazmanian,et al.  Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. , 1999, Science.

[6]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[7]  P. Dawson,et al.  Synthesis of native proteins by chemical ligation. , 2000, Annual review of biochemistry.

[8]  A. Kondo,et al.  Enzyme-mediated site-specific antibody-protein modification using a ZZ domain as a linker. , 2010, Bioconjugate chemistry.

[9]  T. Muir,et al.  Introduction of unnatural amino acids into proteins using expressed protein ligation. , 1999, Biopolymers.

[10]  Hidde L Ploegh,et al.  Sortagging: a versatile method for protein labeling. , 2007, Nature chemical biology.

[11]  H. Ploegh,et al.  Lipid modification of proteins through sortase-catalyzed transpeptidation. , 2008, Journal of the American Chemical Society.

[12]  J. Klose,et al.  Enzymatic ligation of peptides, peptide nucleic acids and proteins by means of sortase A. , 2009, Advances in experimental medicine and biology.

[13]  D. Kojetin,et al.  Observing selected domains in multi-domain proteins via sortase-mediated ligation and NMR spectroscopy , 2011, Journal of biomolecular NMR.

[14]  Ronald T. Raines,et al.  Increasing the secretory capacity of Saccharomyces cerevisiae for production of single-chain antibody fragments , 1998, Nature Biotechnology.

[15]  H. Ploegh,et al.  Sortase-catalyzed transformations that improve the properties of cytokines , 2011, Proceedings of the National Academy of Sciences.

[16]  C. Neylon,et al.  Covalent Attachment of Proteins to Solid Supports and Surfaces via Sortase-Mediated Ligation , 2007, PloS one.

[17]  T. Muir,et al.  Introduction of unnatural amino acids into proteins using expressed protein ligationStandard IUPAC single- and triple-letter codes for amino acids are used throughout the article , 1999 .

[18]  T. Muir,et al.  Protein engineering by expressed protein ligation. , 2000, Methods in enzymology.

[19]  S. Hart,et al.  Sortase-mediated protein ligation: a new method for protein engineering. , 2004, Journal of the American Chemical Society.

[20]  E. Boder,et al.  Sortase A as a novel molecular "stapler" for sequence-specific protein conjugation. , 2007, Bioconjugate chemistry.

[21]  T. Proft,et al.  The use of sortase-mediated ligation for the immobilisation of bacterial adhesins onto fluorescence-labelled microspheres: a novel approach to analyse bacterial adhesion to host cells , 2010, Biotechnology Letters.

[22]  K Dane Wittrup,et al.  A modular IgG-scFv bispecific antibody topology. , 2010, Protein engineering, design & selection : PEDS.

[23]  Teruyuki Nagamune,et al.  Site‐Specific Protein Modification on Living Cells Catalyzed by Sortase , 2008, Chembiochem : a European journal of chemical biology.

[24]  J. Klose,et al.  Synthesis of biologically active peptide nucleic acid-peptide conjugates by sortase-mediated ligation. , 2007, The Journal of organic chemistry.

[25]  T. Proft Sortase-mediated protein ligation: an emerging biotechnology tool for protein modification and immobilisation , 2009, Biotechnology Letters.

[26]  Zhimeng Wu,et al.  Sortase A-catalyzed transpeptidation of glycosylphosphatidylinositol derivatives for chemoenzymatic synthesis of GPI-anchored proteins. , 2010, Journal of the American Chemical Society.

[27]  D. McCafferty,et al.  Mutagenesis Studies of Substrate Recognition and Catalysis in the Sortase A Transpeptidase from Staphylococcus aureus* , 2008, Journal of Biological Chemistry.

[28]  H. Ploegh,et al.  Site‐Specific Protein Labeling via Sortase‐Mediated Transpeptidation , 2009, Current protocols in protein science.

[29]  R. P. Roy,et al.  Peptide-sugar ligation catalyzed by transpeptidase sortase: a facile approach to neoglycoconjugate synthesis. , 2008, Journal of the American Chemical Society.

[30]  T. Proft,et al.  Immobilization of proteins to biacore sensor chips using Staphylococcus aureus sortase A , 2008, Biotechnology Letters.