Amino Acid-Selective Segmental Isotope Labeling of Multidomain Proteins for Structural Biology

ChemBioChem ◽  
2013 ◽  
Vol 14 (4) ◽  
pp. 457-466 ◽  
Author(s):  
Erich Michel ◽  
Lenka Skrisovska ◽  
Kurt Wüthrich ◽  
Frédéric H.-T. Allain
Keyword(s):  
Amino Acid ◽  
Structural Biology ◽  
Isotope Labeling ◽  
Multidomain Proteins ◽  
Segmental Isotope Labeling

Electron probe x-ray microanalysis and electron microscopy of amino acid absorption and transport by the small intestine: inhibition by chemical poisons and correlation to the elemental composition of the epithelial cells

1986 ◽  
Vol 44 ◽  
pp. 134-135
Author(s):  
A. J. Tousimis
Keyword(s):  
Small Intestine ◽  
Amino Acid ◽  
Epithelial Cells ◽  
Elemental Composition ◽  
Isotope Labeling ◽  
Structural Components ◽  
X Ray ◽  
Human Small Intestine ◽  
Transport Studies

The elemental composition of amino acids is similar to that of the major structural components of the epithelial cells of the small intestine and other tissues. Therefore, their subcellular localization and concentration measurements are not possible by x-ray microanalysis. Radioactive isotope labeling: I131-tyrosine, Se75-methionine and S35-methionine have been successfully employed in numerous absorption and transport studies. The latter two have been utilized both in vitro and vivo, with similar results in the hamster and human small intestine. Non-radioactive Selenomethionine, since its absorption/transport behavior is assumed to be the same as that of Se75- methionine and S75-methionine could serve as a compound tracer for this amino acid.

scholarly journals Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling

2005 ◽  
Vol 25 (1) ◽  
pp. 150-162 ◽  
Author(s):  
Francesca Vitali ◽  
Anke Henning ◽  
Florian C Oberstrass ◽  
Yann Hargous ◽  
Sigrid D Auweter ◽  
...  
Keyword(s):  
Isotope Labeling ◽  
Rna Recognition ◽  
Rna Recognition Motifs ◽  
Segmental Isotope Labeling ◽  
Recognition Motifs

scholarly journals Dexamethasone-Induced Perturbations in Tissue Metabolomics Revealed by Chemical Isotope Labeling LC-MS Analysis

Metabolites ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 42 ◽  
Author(s):  
Lina Dahabiyeh ◽  
Abeer Malkawi ◽  
Xiaohang Wang ◽  
Dilek Colak ◽  
Ahmed Mujamammi ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Amino Acid ◽  
Side Effects ◽  
Isotope Labeling ◽  
Protein Biosynthesis ◽  
Chronic Administration ◽  
Therapeutic Interventions ◽  
Sprague Dawley ◽  
The Brain ◽  
Chemical Isotope Labeling

Dexamethasone (Dex) is a synthetic glucocorticoid (GC) drug commonly used clinically for the treatment of several inflammatory and immune-mediated diseases. Despite its broad range of indications, the long-term use of Dex is known to be associated with specific abnormalities in several tissues and organs. In this study, the metabolomic effects on five different organs induced by the chronic administration of Dex in the Sprague–Dawley rat model were investigated using the chemical isotope labeling liquid chromatography-mass spectrometry (CIL LC-MS) platform, which targets the amine/phenol submetabolomes. Compared to controls, a prolonged intake of Dex resulted in significant perturbations in the levels of 492, 442, 300, 186, and 105 metabolites in the brain, skeletal muscle, liver, kidney, and heart tissues, respectively. The positively identified metabolites were mapped to diverse molecular pathways in different organs. In the brain, perturbations in protein biosynthesis, amino acid metabolism, and monoamine neurotransmitter synthesis were identified, while in the heart, pyrimidine metabolism and branched amino acid biosynthesis were the most significantly impaired pathways. In the kidney, several amino acid pathways were dysregulated, which reflected impairments in several biological functions, including gluconeogenesis and ureagenesis. Beta-alanine metabolism and uridine homeostasis were profoundly affected in liver tissues, whereas alterations of glutathione, arginine, glutamine, and nitrogen metabolism pointed to the modulation of muscle metabolism and disturbances in energy production and muscle mass in skeletal muscle. The differential expression of multiple dipeptides was most significant in the liver (down-regulated), brain (up-regulation), and kidney tissues, but not in the heart or skeletal muscle tissues. The identification of clinically relevant pathways provides holistic insights into the tissue molecular responses induced by Dex and understanding of the underlying mechanisms associated with their side effects. Our data suggest a potential role for glutathione supplementation and dipeptide modulators as novel therapeutic interventions to mitigate the side effects induced by Dex therapy.

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.

Segmental isotope-labeling of the intrinsically disordered protein PQBP1

FEBS Letters ◽  
2014 ◽  
Vol 588 (24) ◽  
pp. 4583-4589 ◽  
Author(s):  
Yuko Nabeshima ◽  
Mineyuki Mizuguchi ◽  
Asagi Kajiyama ◽  
Hitoshi Okazawa
Keyword(s):  
Isotope Labeling ◽  
Intrinsically Disordered Protein ◽  
Intrinsically Disordered ◽  
Disordered Protein ◽  
Segmental Isotope Labeling

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 ◽  
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