Cotranslational Incorporation of a Structurally Diverse Series of Proline Analogues in an Escherichia coli Expression System

ChemBioChem ◽  
2004 ◽  
Vol 5 (7) ◽  
pp. 928-936 ◽  
Author(s):  
Wookhyun Kim ◽  
Anna George ◽  
Melissa Evans ◽  
Vincent P. Conticello
Keyword(s):  
Escherichia Coli ◽  
Expression System ◽  
Escherichia Coli Expression

scholarly journals Construction of an antibody Fv-Epo receptor chimera extracellular domainusing Escherichia coli expression system

2000 ◽  
Vol 40 (supplement) ◽  
pp. S186
Author(s):  
Takeshi Kitamura ◽  
Kouhei Tsumoto ◽  
Masahiro Kawahara ◽  
Hiroshi Ueda ◽  
Teruyuki Nagamune ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Expression System ◽  
Epo Receptor ◽  
Escherichia Coli Expression

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.

Effect of rhBMP-2 produced by Escherichia coli expression system on bone formation in rat calvarial defects

2009 ◽  
Vol 39 (1) ◽  
pp. 77
Author(s):  
Suk-Hoon Kwon ◽  
Hyun-Chang Lim ◽  
Kyung-Hee Choi ◽  
Min-Soo Kim ◽  
Ji-Hyun Lee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bone Formation ◽  
Expression System ◽  
Calvarial Defects ◽  
Escherichia Coli Expression

scholarly journals Production of an authentic-like recombinant protein based on Escherichia coli expression system

2000 ◽  
Vol 40 (supplement) ◽  
pp. S185
Author(s):  
T. Koshiba ◽  
T. Tamada ◽  
K. Nitta ◽  
R. Kuroki
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Expression System ◽  
Escherichia Coli Expression

scholarly journals Co-expression of glutathione S-transferase with methionine aminopeptidase: a system of producing enriched N-terminal processed proteins in Escherichia coli

1999 ◽  
Vol 338 (2) ◽  
pp. 335-342 ◽  
Author(s):  
Debbie D. W. HWANG ◽  
Li-Fan LIU ◽  
I-Ching KUAN ◽  
Lih-Yuan LIN ◽  
Tsuey-Chyi S. TAM ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Expression System ◽  
Methionine Aminopeptidase ◽  
Glutathione S Transferase ◽  
Methionine Residue ◽  
Methionine Residues ◽  
Processed Form ◽  
Escherichia Coli Expression ◽  
Rat Livers

We describe here an Escherichia coli expression system that produces recombinant proteins enriched in the N-terminal processed form, by using glutathione S-transferase cGSTM1-1 and rGSTT1-1 as models, where c and r refer to chick and rat respectively. Approximately 90% of the cGSTM1-1 or rGSTT1-1 overexpressed in E. coliunder the control of a phoA promoter retained the initiator methionine residue that was absent from the mature isoenzymes isolated from tissues. The amount of initiator methionine was decreased to 40% of the expressed cGSTM1-1 when the isoenzyme was co-expressed with an exogenous methionine aminopeptidase gene under the control of a separate phoA promoter. The recombinant proteins expressed were mainly methionine aminopeptidase. The yield of cGSTM1-1 was decreased to 10% of that expressed in the absence of the exogenous methionine aminopeptidase gene. By replacing the phoA with its natural promoter, the expression of methionine aminopeptidase decreased drastically. The yield of the co-expressed cGSTM1-1 was approx. 60% of that in the absence of the exogenous methionine aminopeptidase gene; approx. 65% of the initiator methionine residues were removed from the enzyme. Under similar conditions, N-terminal processing was observed in approx. 70% of the recombinant rGSTT1-1 expressed. By increasing the concentration of phosphate in the growth medium, the amount of initiator methionine on cGSTM1-1 was decreased to 14% of the overexpressed isoenzymes, whereas no further improvement could be observed for rGSTT1-1. The initiator methionine residue does not affect the enzymic activities of either cGSTM1-1 or rGSTT1-1. However, the epoxidase activity and the 4-nitrobenzyl chloride-conjugating activity of the purified recombinant rGSTT1-1 are markedly higher that those reported recently for the same isoenzyme isolated from rat livers.

Sign in / Sign up

Export Citation Format

Share Document