Site-specific labeling of DNA–protein conjugates by means of expressed protein ligation

2007 ◽  
pp. 353-355 ◽  
Author(s):  
Marina Lovrinovic ◽  
Ljiljana Fruk ◽  
Hendrik Schröder ◽  
Christof M. Niemeyer
Keyword(s):  
Expressed Protein Ligation ◽  
Site Specific ◽  
Protein Ligation ◽  
Protein Conjugates

scholarly journals Analysis of Site‐Specific Phosphorylation of PTEN by Using Enzyme‐Catalyzed Expressed Protein Ligation

ChemBioChem ◽  
2019 ◽  
Vol 21 (1-2) ◽  
pp. 64-68 ◽  
Author(s):  
Samuel H. Henager ◽  
Stephanie Henriquez ◽  
Daniel R. Dempsey ◽  
Philip A. Cole
Keyword(s):  
Expressed Protein Ligation ◽  
Site Specific ◽  
Protein Ligation

scholarly journals Sortase-Tag Expressed Protein Ligation: Combining Protein Purification and Site-Specific Bioconjugation into a Single Step

2013 ◽  
Vol 85 (22) ◽  
pp. 11090-11097 ◽  
Author(s):  
Robert Warden-Rothman ◽  
Ilaria Caturegli ◽  
Vladimir Popik ◽  
Andrew Tsourkas
Keyword(s):  
Protein Purification ◽  
Single Step ◽  
Expressed Protein Ligation ◽  
Site Specific ◽  
Protein Ligation

Site-Specific Antibody Drug Conjugates Using Streamlined Expressed Protein Ligation

2018 ◽  
Vol 29 (11) ◽  
pp. 3503-3508 ◽  
Author(s):  
Silvia Frutos ◽  
Jose Luis Hernández ◽  
Anabel Otero ◽  
Carme Calvis ◽  
Jaume Adan ◽  
...  
Keyword(s):  
Specific Antibody ◽  
Expressed Protein Ligation ◽  
Site Specific ◽  
Antibody Drug Conjugates ◽  
Protein Ligation ◽  
Drug Conjugates ◽  
Antibody Drug

Synthesis of covalent DNA–protein conjugates by expressed protein ligation

2005 ◽  
Vol 1 (1) ◽  
pp. 64 ◽  
Author(s):  
Marina Lovrinovic ◽  
Mark Spengler ◽  
Carl Deutsch ◽  
Christof M. Niemeyer
Keyword(s):  
Expressed Protein Ligation ◽  
Protein Ligation ◽  
Protein Conjugates

scholarly journals Site-specific conjugation of RAFT polymers to proteins via expressed protein ligation

2013 ◽  
Vol 49 (25) ◽  
pp. 2566 ◽  
Author(s):  
Yan Xia ◽  
Shengchang Tang ◽  
Bradley D. Olsen
Keyword(s):  
Expressed Protein Ligation ◽  
Site Specific ◽  
Protein Ligation ◽  
Raft Polymers

scholarly journals Expressed Protein Ligation Without Intein

2020 ◽  
Author(s):  
Yuchen Qiao ◽  
Ge Yu ◽  
Xiaoyan Wang ◽  
Kaci C. Kratch ◽  
Wesley Wei Wang ◽  
...  
Keyword(s):  
Rnase H ◽  
Protein Chemistry ◽  
Histone H2a ◽  
Expressed Protein Ligation ◽  
Protein Ligation ◽  
Protein Conjugates ◽  
Synthetic Capacity ◽  
New Research ◽  
Nucleophilic Acyl Substitution ◽  
Fusion Approach

Proteins with a functionalized <i>C</i>-terminus such as a <i>C</i>-terminal thioester are key to the synthesis of larger proteins via expressed protein ligation. They are usually made by recombinant fusion to intein. Although powerful, the intein fusion approach suffers from premature hydrolysis and low compatibility with denatured conditions. To totally bypass the involvement of an enzyme for expressed protein ligation, here we showed that a cysteine in a recombinant protein was chemically activated by a small molecule cyanylating reagent at its <i>N</i>-side amide for undergoing nucleophilic acyl substitution with amines including a number of L- and D-amino acids and hydrazine. The afforded protein hydrazides could be used further for expressed protein ligation. We demonstrated the versatility of this approach with the successful synthesis of ubiquitin conjugates, ubiquitin-like protein conjugates, histone H2A with a posttranslational modification, RNAse H that actively hydrolyzed RNA, and exenatide that is a commercial therapeutic peptide. The technique, which is exceedingly simple but highly useful, expands to a great extent the synthetic capacity of protein chemistry and will therefore make a large avenue of new research possible.

scholarly journals Access to site-specific Fc–cRGD peptide conjugates through streamlined expressed protein ligation

2016 ◽  
Vol 14 (40) ◽  
pp. 9549-9553 ◽  
Author(s):  
S. Frutos ◽  
J. B. Jordan ◽  
M. M. Bio ◽  
T. W. Muir ◽  
O. R. Thiel ◽  
...  
Keyword(s):  
Active Site ◽  
Bioactive Peptides ◽  
Expressed Protein Ligation ◽  
Peptide Conjugates ◽  
Site Specific ◽  
Protein Ligation ◽  
Split Intein ◽  
Efficient Route ◽  
Long Acting ◽  
Powerful Strategy

Conjugation of peptides to the Fc fragment of antibodies is a powerful strategy to generate long acting biotherapeutics. We show here an efficient route to obtain fully active, site-specific conjugates of synthetic bioactive peptides using a split intein based approach.

scholarly journals Expressed Protein Ligation Without Intein

2020 ◽  
Author(s):  
Yuchen Qiao ◽  
Ge Yu ◽  
Xiaoyan Wang ◽  
Kaci C. Kratch ◽  
Wesley Wei Wang ◽  
...  
Keyword(s):  
Rnase H ◽  
Protein Chemistry ◽  
Histone H2a ◽  
Expressed Protein Ligation ◽  
Protein Ligation ◽  
Protein Conjugates ◽  
Synthetic Capacity ◽  
New Research ◽  
Nucleophilic Acyl Substitution ◽  
Fusion Approach

Proteins with a functionalized <i>C</i>-terminus such as a <i>C</i>-terminal thioester are key to the synthesis of larger proteins via expressed protein ligation. They are usually made by recombinant fusion to intein. Although powerful, the intein fusion approach suffers from premature hydrolysis and low compatibility with denatured conditions. To totally bypass the involvement of an enzyme for expressed protein ligation, here we showed that a cysteine in a recombinant protein was chemically activated by a small molecule cyanylating reagent at its <i>N</i>-side amide for undergoing nucleophilic acyl substitution with amines including a number of L- and D-amino acids and hydrazine. The afforded protein hydrazides could be used further for expressed protein ligation. We demonstrated the versatility of this approach with the successful synthesis of ubiquitin conjugates, ubiquitin-like protein conjugates, histone H2A with a posttranslational modification, RNAse H that actively hydrolyzed RNA, and exenatide that is a commercial therapeutic peptide. The technique, which is exceedingly simple but highly useful, expands to a great extent the synthetic capacity of protein chemistry and will therefore make a large avenue of new research possible.

Sign in / Sign up

Export Citation Format

Share Document