scholarly journals Enlightening advances in polymer bioconjugate chemistry: light-based techniques for grafting to and from biomacromolecules

2020 ◽  
Vol 11 (20) ◽  
pp. 5142-5156 ◽  
Author(s):  
Rebecca A. Olson ◽  
Angie B. Korpusik ◽  
Brent S. Sumerlin
Keyword(s):  
Protein Ligation ◽  
Bioconjugate Chemistry

Light-driven synthetic techniques have revolutionized polymer–biomacromolecule conjugation. This Perspective discusses the benefits of photochemistry for polymer–protein ligation and considerations when designing a new light-driven conjugation approach.

Enhancement of sortase A-mediated protein ligation by inducing a β-hairpin structure around the ligation site

2011 ◽  
Vol 47 (16) ◽  
pp. 4742 ◽  
Author(s):  
Yuichi Yamamura ◽  
Hidehiko Hirakawa ◽  
Satoshi Yamaguchi ◽  
Teruyuki Nagamune
Keyword(s):  
Hairpin Structure ◽  
Sortase A ◽  
Protein Ligation ◽  
Ligation Site

scholarly journals Simple bioconjugate chemistry serves great clinical advances: albumin as a versatile platform for diagnosis and precision therapy

2016 ◽  
Vol 45 (5) ◽  
pp. 1432-1456 ◽  
Author(s):  
Zhibo Liu ◽  
Xiaoyuan Chen
Keyword(s):  
Drug Targeting ◽  
Pharmacokinetic Profile ◽  
Protein Carrier ◽  
Precision Therapy ◽  
Bioconjugate Chemistry ◽  
Clinical Advances

Albumin is the most abundant circulating protein in plasma and has recently emerged as a versatile protein carrier for drug targeting and for improving the pharmacokinetic profile of peptide or protein based drugs.

scholarly journals In Vivo and In Vitro Protein Ligation by Naturally Occurring and Engineered Split DnaE Inteins

PLoS ONE ◽  
2009 ◽  
Vol 4 (4) ◽  
pp. e5185 ◽  
Author(s):  
A. Sesilja Aranko ◽  
Sara Züger ◽  
Edith Buchinger ◽  
Hideo Iwaï
Keyword(s):  
Protein Ligation ◽  
Naturally Occurring ◽  
Vitro Protein

scholarly journals Analysis of Protein Kinase Autophosphorylation Using Expressed Protein Ligation and Computational Modeling

2008 ◽  
Vol 130 (17) ◽  
pp. 5667-5669 ◽  
Author(s):  
Kerry A. Pickin ◽  
Sidhartha Chaudhury ◽  
Blair C. R. Dancy ◽  
Jeffrey J. Gray ◽  
Philip A. Cole
Keyword(s):  
Protein Kinase ◽  
Computational Modeling ◽  
Expressed Protein Ligation ◽  
Protein Ligation

scholarly journals Display of Recombinant Proteins on Bacterial Outer Membrane Vesicles by Using Protein Ligation

2018 ◽  
Vol 84 (8) ◽  
pp. e02567-17 ◽  
Author(s):  
H. Bart van den Berg van Saparoea ◽  
Diane Houben ◽  
Marien I. de Jonge ◽  
Wouter S. P. Jong ◽  
Joen Luirink
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Salmonella Enterica ◽  
Membrane Vesicles ◽  
Therapeutic Agents ◽  
Outer Membrane Vesicles ◽  
High Density ◽  
Content Type ◽  
Protein Ligation ◽  
Serovar Typhimurium

ABSTRACT The Escherichia coli virulence factor hemoglobin protease (Hbp) has been engineered into a surface display system that can be expressed to high density on live E. coli and Salmonella enterica serovar Typhimurium cells or derived outer membrane vesicles (OMVs). Multiple antigenic sequences can be genetically fused into the Hbp core structure for optimal exposure to the immune system. Although the Hbp display platform is relatively tolerant, increasing the number, size, and complexity of integrated sequences generally lowers the expression of the fused constructs and limits the density of display. This is due to the intricate mechanism of Hbp secretion across the outer membrane and the efficient quality control of translocation-incompetent chimeric Hbp molecules in the periplasm. To address this shortcoming, we explored the coupling of purified proteins to the Hbp carrier after its translocation across the outer membrane using the recently developed SpyTag/SpyCatcher protein ligation system. As expected, fusion of the small SpyTag to Hbp did not hamper display on OMVs. Subsequent addition of purified proteins fused to the SpyCatcher domain resulted in efficient covalent coupling to Hbp-SpyTag. Using in addition the orthogonal SnoopTag/SnoopCatcher system, multiple antigen modules could be coupled to Hbp in a sequential ligation strategy. Not only antigens proved suitable for Spy-mediated ligation but also nanobodies. Addition of this functionality to the platform might allow the targeting of live bacterial or OMV vaccines to certain tissues or immune cells to tailor immune responses.IMPORTANCE Outer membrane vesicles (OMVs) derived from Gram-negative bacteria attract increasing interest in the development of vaccines and therapeutic agents. We aim to construct a semisynthetic OMV platform for recombinant antigen presentation on OMVs derived from attenuated Salmonella enterica serovar Typhimurium cells displaying an adapted Escherichia coli autotransporter, Hbp, at the surface. Although this autotransporter accepts substantial modifications, its capacity with respect to the number, size, and structural complexity of the antigens genetically fused to the Hbp carrier is restricted. Here we describe the application of SpyCatcher/SpyTag protein ligation technology to enzymatically link antigens to Hbp present at high density in OMVs. Protein ligation was apparently unobstructed by the membrane environment and allowed a high surface density of coupled antigens, a property we have shown to be important for vaccine efficacy. The OMV coupling procedure appears versatile and robust, allowing fast production of experimental vaccines and therapeutic agents through a modular plug-and-display procedure.

Sign in / Sign up

Export Citation Format

Share Document