One-step amino acid selective isotope labeling of proteins in prototrophic Escherichia coli strains

2012 ◽  
Vol 426 (2) ◽  
pp. 126-128 ◽  
Author(s):  
Christopher O’Grady ◽  
Benjamin L. Rempel ◽  
Akosiererem Sokaribo ◽  
Sergiy Nokhrin ◽  
Oleg Y. Dmitriev
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Isotope Labeling ◽  
Selective Isotope Labeling ◽  
One Step

scholarly journals Escherichia coli amino acid auxotrophic expression host strains for investigating protein structure-function relationships

2020 ◽  
Author(s):  
Toshio Iwasaki ◽  
Yoshiharu Miyajima-Nakano ◽  
Risako Fukazawa ◽  
Myat T Lin ◽  
Shin-Ichi Matsushita ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Structure ◽  
Amino Acid ◽  
Structure Function ◽  
Growth Medium ◽  
Isotope Labeling ◽  
Water Soluble ◽  
Expression Host ◽  
Host Strains

Abstract A set of C43(DE3) and BL21(DE3) Escherichia coli host strains that are auxotrophic for various amino acids is briefly reviewed. These strains require the addition of a defined set of one or more amino acids in the growth medium, and have been specifically designed for overproduction of membrane or water-soluble proteins selectively labeled with stable isotopes such as 2H, 13C and 15N. The strains described here are available for use and have been deposited into public strain banks. Although they cannot fully eliminate the possibility of isotope dilution and mixing, metabolic scrambling of the different amino acid types can be minimized through a careful consideration of the bacterial metabolic pathways. The use of a suitable auxotrophic expression host strain with an appropriately isotopically labeled growth medium ensures high levels of isotope labeling efficiency as well as selectivity for providing deeper insight into protein structure-function relationships.

A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed inLeishmania tarentolae

2009 ◽  
Vol 26 (6) ◽  
pp. 755-761 ◽  
Author(s):  
Silvie Foldynová-Trantírková ◽  
Jana Matulová ◽  
Volker Dötsch ◽  
Frank Löhr ◽  
Ion Cirstea ◽  
...  
Keyword(s):  
Amino Acid ◽  
Isotope Labeling ◽  
Cost Effective ◽  
Amino Acid Type ◽  
Acid Type ◽  
Selective Isotope Labeling

scholarly journals Cloning, sequencing and overexpression in Escherichia coli of the alginatelyase-encoding aly gene of Pseudomonas alginovora: identification of three classes of alginate lyases

1996 ◽  
Vol 319 (2) ◽  
pp. 575-583 ◽  
Author(s):  
Frederic CHAVAGNAT ◽  
Colette DUEZ ◽  
Micheline GUINAND ◽  
Philippe POTIN ◽  
Tristan BARBEYRON ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Alginate Lyase ◽  
Wild Type ◽  
E Coli ◽  
Step Procedure ◽  
Amino Acid Precursor ◽  
One Step ◽  
Alginate Lyases ◽  
Pcr Techniques

A gene of Pseudomonas alginovora, called aly, has been cloned in Escherichia coli using a battery of PCR techniques and sequenced. It encodes a 210-amino-acid alginate lyase (EC 4.2.2.3), Aly, in the form of a 233-amino-acid precursor. P. alginovora Aly has been overproduced in E. coli with a His-tag sequence fused at the C-terminal end under conditions in which the formation of inclusion bodies is avoided. His-tagged P. alginovora Aly has the same enzymic properties as the wild-type enzyme and has the specificity of a mannuronate lyase. It can be purified in a one-step procedure by affinity chromatography on Ni2+-nitriloacetate resin. The yield is of 5 mg of enzyme per litre of culture. The amplification factor is 12.5 compared with the level of production by wild-type P. alginovora. The six alginate lyases of known primary structure fall into three distinct classes, one of which comprises the pair P. alginovora Aly and Klebsiella pneumoniae Aly.

scholarly journals Amino-acid selective isotope labeling enables simultaneous overlapping signal decomposition and information extraction from NMR spectra

2020 ◽  
Vol 74 (2-3) ◽  
pp. 125-137
Author(s):  
Takuma Kasai ◽  
Shunsuke Ono ◽  
Seizo Koshiba ◽  
Masayuki Yamamoto ◽  
Toshiyuki Tanaka ◽  
...  
Keyword(s):  
Amino Acid ◽  
Information Extraction ◽  
Nmr Spectra ◽  
Isotope Labeling ◽  
Signal Decomposition ◽  
Selective Isotope Labeling

A simple protocol for amino acid type selective isotope labeling in insect cells with improved yields and high reproducibility

2011 ◽  
Vol 51 (4) ◽  
pp. 449-456 ◽  
Author(s):  
Alvar D. Gossert ◽  
Alexandra Hinniger ◽  
Sascha Gutmann ◽  
Wolfgang Jahnke ◽  
André Strauss ◽  
...  
Keyword(s):  
Amino Acid ◽  
Insect Cells ◽  
Isotope Labeling ◽  
Amino Acid Type ◽  
High Reproducibility ◽  
Acid Type ◽  
Simple Protocol ◽  
Selective Isotope Labeling
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