Total Chemical Synthesis of the Enzyme Sortase AΔN59with Full Catalytic Activity

2014 ◽  
Vol 53 (18) ◽  
pp. 4662-4666 ◽  
Author(s):  
Fang-Kun Deng ◽  
Liang Zhang ◽  
Ya-Ting Wang ◽  
Olaf Schneewind ◽  
Stephen B. H. Kent
Keyword(s):  
Catalytic Activity ◽  
Chemical Synthesis

Expression of adenylate kinase fused mouse ubiquitin active enzyme in Escherichia coli and its application in ubiquitination

2021 ◽  
Author(s):  
Xiaoliang Liu ◽  
Ling Hu ◽  
Yuan Zhang ◽  
Hongtao Li
Keyword(s):  
Escherichia Coli ◽  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Adenylate Kinase ◽  
Energy Supply ◽  
Expression System ◽  
Synthesis Process ◽  
Cellular Functions ◽  
Escherichia Coli Expression

Abstract Ubiquitination, is involved in the regulation of numerous cellular functions. Researches in the ubiquitin realm rely heavily on ubiquitination assays in vitro and require large amounts of ubiquitin-activating enzyme (UBA1) and keep ATP supplies. But UBA1 is hard to be obtained with large quantities using reported methods. We fused Escherichia coli adenylate kinase (adk) and mouse UBA1 obtained fusion protein adk-mUBA1. The expression level of adk-mUBA1 increased about 8-fold than that of mUBA1 in Escherichia coli expression system, and adk-mUBA1 was easily purified to 90% purity via two purification steps. The purified adk-mUBA1 protein was functional for ubiquitination and could use ATP in addition to ADP as energy supply and had a higher catalytic activity than mUBA1 in cell lysis. Adk-mUBA1 can be applied to preparing ubiquitin modified substrates and kinds of ubiquitin chains in chemical synthesis process and is preferable application than mUBA1 in vitro ubiquitination.

scholarly journals Copper-mediated selenazolidine deprotection enables one-pot chemical synthesis of challenging proteins

2019 ◽  
Author(s):  
Zhenguang Zhao ◽  
Norman Metanis
Keyword(s):  
Protein Synthesis ◽  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Total Synthesis ◽  
Active Site ◽  
Synthetic Approach ◽  
One Pot ◽  
Chemical Protein Synthesis ◽  
Key Technologies ◽  
The One

<p>While chemical protein synthesis (CPS) has granted access to challenging proteins, synthesis of longer proteins is often limited by low abundance or non-strategic placement of cysteine (Cys) residues, essential for native chemical ligations (NCL), as well as multiple purification and isolation steps. Selective deselenization and one-pot CPS serve as key technologies to circumvent these issues. Herein, we describe the one-pot total synthesis of human thiosulfate: glutathione sulfurtransferase (TSTD1), a 115-residue protein with a single Cys residue at its active site, and its seleno-analogue. WT-TSTD1 was synthesized in a C-to-N synthetic approach employing multiple NCL reactions, Cu(II)-mediated deprotection of selenazolidine (Sez), and chemoselective deselenization, all in one-pot. In addition, the protein’s seleno analogue (Se-TSTD1), in which the active site Cys is replaced with selenocysteine, was synthesized with a kinetically controlled ligation in a one-pot, N-to-C synthetic approach. TSTD1’s one-pot synthesis was made possible by the newly reported, rapid, and facile copper-mediated selenazolidine deprotection that can be accomplished in one minute. Finally, catalytic activity of the two proteins indicated that Se-TSTD1 possessed only four-fold lower activity than WT-TSTD1 as a thiosulfate: glutathione sulfurtransferase, suggesting that selenoproteins can have physiologically comparable sulfutransferase activity as their cysteine counterparts. </p>

One-step, seedless wet-chemical synthesis of gold@palladium nanoflowers supported on reduced graphene oxide with enhanced electrocatalytic properties

2014 ◽  
Vol 2 (43) ◽  
pp. 18177-18183 ◽  
Author(s):  
Shan-Shan Li ◽  
Ai-Jun Wang ◽  
Yuan-Yuan Hu ◽  
Ke-Ming Fang ◽  
Jian-Rong Chen ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Alkaline Media ◽  
Chemical Route ◽  
Wet Chemical Synthesis ◽  
Reduced Graphene ◽  
One Step ◽  
Wet Chemical ◽  
Wet Chemical Route

A simple, seedless wet-chemical route was designed for synthesis of well-dispersed Au@Pd nanoflowers on rGO. The as-prepared nanocomposites exhibited enhanced catalytic activity and better stability for ORR and EG oxidation in alkaline media.

Chemical synthesis and enzymatic properties of RNase A analogues designed to enhance second-step catalytic activity

2016 ◽  
Vol 14 (37) ◽  
pp. 8804-8814
Author(s):  
David J. Boerema ◽  
Valentina A. Tereshko ◽  
Junliang Zhang ◽  
Stephen B. H. Kent
Keyword(s):  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Rnase A ◽  
Enzymatic Properties ◽  
Second Step ◽  
Enzyme Molecule ◽  
Rate Of Hydrolysis

Adenine covalently attached to the RNase A enzyme molecule decreased the rate of transphosphorylation and increased the rate of hydrolysis.

scholarly journals Copper-mediated selenazolidine deprotection enables one-pot chemical synthesis of challenging proteins

2019 ◽  
Author(s):  
Zhenguang Zhao ◽  
Norman Metanis
Keyword(s):  
Protein Synthesis ◽  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Total Synthesis ◽  
Active Site ◽  
Synthetic Approach ◽  
One Pot ◽  
Chemical Protein Synthesis ◽  
Key Technologies ◽  
The One

<p>While chemical protein synthesis (CPS) has granted access to challenging proteins, synthesis of longer proteins is often limited by low abundance or non-strategic placement of cysteine (Cys) residues, essential for native chemical ligations (NCL), as well as multiple purification and isolation steps. Selective deselenization and one-pot CPS serve as key technologies to circumvent these issues. Herein, we describe the one-pot total synthesis of human thiosulfate: glutathione sulfurtransferase (TSTD1), a 115-residue protein with a single Cys residue at its active site, and its seleno-analogue. WT-TSTD1 was synthesized in a C-to-N synthetic approach employing multiple NCL reactions, Cu(II)-mediated deprotection of selenazolidine (Sez), and chemoselective deselenization, all in one-pot. In addition, the protein’s seleno analogue (Se-TSTD1), in which the active site Cys is replaced with selenocysteine, was synthesized with a kinetically controlled ligation in a one-pot, N-to-C synthetic approach. TSTD1’s one-pot synthesis was made possible by the newly reported, rapid, and facile copper-mediated selenazolidine deprotection that can be accomplished in one minute. Finally, catalytic activity of the two proteins indicated that Se-TSTD1 possessed only four-fold lower activity than WT-TSTD1 as a thiosulfate: glutathione sulfurtransferase, suggesting that selenoproteins can have physiologically comparable sulfutransferase activity as their cysteine counterparts. </p>

Chemical Synthesis of an Enzyme Containing an Artificial Catalytic Apparatus

2020 ◽  
Vol 73 (4) ◽  
pp. 321
Author(s):  
Vladimir Torbeev ◽  
Stephen B. H. Kent
Keyword(s):  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Enzyme Catalysis ◽  
Polypeptide Chain ◽  
Peptide Bond ◽  
Enzyme Molecule ◽  
Peptide Substrates ◽  
Unique Approach ◽  
Hiv 1 ◽  
Residue Polypeptide

With the goal of investigating electronic aspects of the catalysis of peptide bond hydrolysis, an analogue of HIV-1 protease was designed in which a non-peptide hydroxy-isoquinolinone artificial catalytic apparatus replaced the conserved Asp25–Thr26–Gly27 sequence in each 99-residue polypeptide chain of the homodimeric enzyme molecule. The enzyme analogue was prepared by total chemical synthesis and had detectable catalytic activity on known HIV-1 protease peptide substrates. Compared with uncatalyzed hydrolysis, the analogue enzyme increased the rate of peptide bond hydrolysis by ∼108-fold. Extensions of this unique approach to the study of enzyme catalysis in HIV-1 protease are discussed.

scholarly journals Highly porous palladium nanodendrites: wet-chemical synthesis, electron tomography and catalytic activity

2019 ◽  
Vol 48 (11) ◽  
pp. 3758-3767 ◽  
Author(s):  
Stefanos Mourdikoudis ◽  
Verónica Montes-García ◽  
Sergio Rodal-Cedeira ◽  
Naomi Winckelmans ◽  
Ignacio Pérez-Juste ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Chemical Synthesis ◽  
Electron Tomography ◽  
Simple Procedure ◽  
Wet Chemical Synthesis ◽  
One Step ◽  
Wet Chemical ◽  
Highly Porous

A simple procedure to obtain highly porous hydrophilic palladium nanodendrites in one step is described.

Total Chemical Synthesis of the Enzyme Sortase AΔN59with Full Catalytic Activity

2014 ◽  
Vol 126 (18) ◽  
pp. 4750-4754 ◽  
Author(s):  
Fang-Kun Deng ◽  
Liang Zhang ◽  
Ya-Ting Wang ◽  
Olaf Schneewind ◽  
Stephen B. H. Kent
Keyword(s):  
Catalytic Activity ◽  
Chemical Synthesis
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