scholarly journals Broad substrate tolerance of tubulin tyrosine ligase enables one-step site-specific enzymatic protein labeling

2017 ◽  
Vol 8 (5) ◽  
pp. 3471-3478 ◽  
Author(s):  
Dominik Schumacher ◽  
Oliver Lemke ◽  
Jonas Helma ◽  
Lena Gerszonowicz ◽  
Verena Waller ◽  
...  
Keyword(s):  
Protein Labeling ◽  
Site Specific ◽  
Wide Applicability ◽  
One Step ◽  
Tubulin Tyrosine Ligase ◽  
Protein Functionalization ◽  
Substrate Tolerance ◽  
Step Site

The broad substrate tolerance of tubulin tyrosine ligase enables its wide applicability for protein functionalization.

One-step site-specific antibody fragment auto-conjugation using SNAP-tag technology

2019 ◽  
Vol 14 (11) ◽  
pp. 3101-3125 ◽  
Author(s):  
Ahmad Fawzi Hussain ◽  
Paul A. Heppenstall ◽  
Florian Kampmeier ◽  
Ivo Meinhold-Heerlein ◽  
Stefan Barth
Keyword(s):  
Specific Antibody ◽  
Antibody Fragment ◽  
Site Specific ◽  
One Step ◽  
Step Site

Lysine Acylation Using Conjugating Enzymes (LACE) for Site-Specific Modification and Ubiquitination of Native Proteins

2019 ◽  
Author(s):  
Raphael Hofmann ◽  
Gaku Akimoto ◽  
Thomas G. Wucherpfennig ◽  
Cathleen Zeymer ◽  
Jeffrey Bode
Keyword(s):  
Protein Labeling ◽  
Sequence Specificity ◽  
Reaction Conditions ◽  
One Pot ◽  
Site Specific ◽  
Protein Substrates ◽  
Ubiquitinated Proteins ◽  
Native Proteins ◽  
Lysine Residues ◽  
Substrate Tolerance

<p>Enzymes are powerful tools for post-translational protein labeling due to their high sequence specificity and mild reaction conditions. Many existing protocols, however, are restricted to conjugations at terminal positions or rely on non-peptidic metabolites and large recognition domains. Here we introduce a chemoenzymatic method to functionalize proteins at internal lysine residues that are part of genetically encoded minimal recognition tags (four residues). We achieved this by employing the intrinsic sequence specificity of the E2 SUMO-conjugating enzyme Ubc9 and a short peptide thioester, which together obviate the need for E1 and E3 enzymes. Using a range of protein substrates, we apply this approach to the conjugation of biochemical probes, one-pot dual-labeling reactions in combination with sortase, and site-specific monoubiquitination and ISG15ylation. The small tag size and large substrate tolerance of Ubc9 will make this a method of choice for protein engineering by isopeptide formation and the preparation of ubiquitinated proteins. </p>

One-Step Fluorescent Protein Labeling by Tubulin Tyrosine Ligase

2019 ◽  
pp. 167-189
Author(s):  
Dominik Schumacher ◽  
Heinrich Leonhardt ◽  
Christian P. R. Hackenberger ◽  
Jonas Helma
Keyword(s):  
Fluorescent Protein ◽  
Protein Labeling ◽  
One Step ◽  
Tubulin Tyrosine Ligase

One-step site-specific modification of native proteins with 2-pyridinecarboxyaldehydes

2015 ◽  
Vol 11 (5) ◽  
pp. 326-331 ◽  
Author(s):  
James I MacDonald ◽  
Henrik K Munch ◽  
Troy Moore ◽  
Matthew B Francis
Keyword(s):  
Site Specific ◽  
Specific Modification ◽  
Native Proteins ◽  
One Step ◽  
Step Site

Lysine Acylation Using Conjugating Enzymes (LACE) for Site-Specific Modification and Ubiquitination of Native Proteins

2019 ◽  
Author(s):  
Raphael Hofmann ◽  
Gaku Akimoto ◽  
Thomas G. Wucherpfennig ◽  
Cathleen Zeymer ◽  
Jeffrey Bode
Keyword(s):  
Protein Labeling ◽  
Sequence Specificity ◽  
Reaction Conditions ◽  
One Pot ◽  
Site Specific ◽  
Protein Substrates ◽  
Ubiquitinated Proteins ◽  
Native Proteins ◽  
Lysine Residues ◽  
Substrate Tolerance

<p>Enzymes are powerful tools for post-translational protein labeling due to their high sequence specificity and mild reaction conditions. Many existing protocols, however, are restricted to conjugations at terminal positions or rely on non-peptidic metabolites and large recognition domains. Here we introduce a chemoenzymatic method to functionalize proteins at internal lysine residues that are part of genetically encoded minimal recognition tags (four residues). We achieved this by employing the intrinsic sequence specificity of the E2 SUMO-conjugating enzyme Ubc9 and a short peptide thioester, which together obviate the need for E1 and E3 enzymes. Using a range of protein substrates, we apply this approach to the conjugation of biochemical probes, one-pot dual-labeling reactions in combination with sortase, and site-specific monoubiquitination and ISG15ylation. The small tag size and large substrate tolerance of Ubc9 will make this a method of choice for protein engineering by isopeptide formation and the preparation of ubiquitinated proteins. </p>

Versatile and Efficient Site-Specific Protein Functionalization by Tubulin Tyrosine Ligase

2015 ◽  
Vol 54 (46) ◽  
pp. 13787-13791 ◽  
Author(s):  
Dominik Schumacher ◽  
Jonas Helma ◽  
Florian A. Mann ◽  
Garwin Pichler ◽  
Francesco Natale ◽  
...  
Keyword(s):  
Specific Protein ◽  
Site Specific ◽  
Tubulin Tyrosine Ligase ◽  
Protein Functionalization

Selective Functionalization at N2-Position of Guanine in Oligonucleotides via Reductive Amination

2020 ◽  
Author(s):  
Bapurao Bhoge ◽  
Ishu Saraogi
Keyword(s):  
Organic Chemistry ◽  
Chemical Biology ◽  
Reductive Amination ◽  
The Other ◽  
Dna Oligomers ◽  
Site Specific ◽  
Wide Applicability ◽  
Dna Oligonucleotides ◽  
Selective Functionalization

Chemo- and site-specific modifications in oligonucleotides have wide applicability as mechanistic probes in chemical biology. Here we have employed a classical reaction in organic chemistry, reductive amination, to selectively functionalize the N<sup>2</sup>-amine of guanine/2’-deoxyguanine monophosphate. This method specifically modifies guanine in several tested DNA oligonucleotides, while leaving the other bases unaffected. Using this approach, we have successfully incorporated desired handles chemoselectively into DNA oligomers.

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