Ergothioneine is an emerging component of the redox homeostasis system in human cells and microbial pathogens, such as Mycobacterium tuberculosis Burkholderia pseudomallei. The synthesis stable isotope-labeled ergothioneine derivatives may provide important tools for deciphering distribution, function, metabolism this compound vivo. We describe a general protocol production isotopologues with p...

Protein turnover in living systems has been measured with the use of stable isotope labeled tracers for over half a century [1]. Recent advances, in mass spectrometry, sample preparation and separation sciences have made it possible for this approach to become applicable at a global proteomics level, permitting analysis of the turnover of many proteins instead of single proteins or an aggregate...

A rapid and efficient approach for preparing isotopically labeled recombinant proteins is presented. The method is demonstrated for 13C labeling of the C-terminal domain of angiopoietin-2, 15N labeling of ubiquitin and for 2H/13C/15N labeling of the Escherichia coli outer-membrane lipoprotein Lpp-56. The production method generates cell mass using unlabeled rich media followed by exchange into ...

Cell cultures of Catharanthus roseus were supplied with [2-13C, 3-2H]-deoxyxylulose or [2-13C,4-2H]1-deoxyxylulose. Lutein and chlorophylls were isolated from the cell mass, and hydrolysis of the chlorophyll mixtures afforded phytol. Isotope labeling patterns of phytol and lutein were determined by 2H NMR and 1H,2H-decoupled 13C NMR. From the data it must be concluded that the deuterium atom in...

Deuterium substitution on two ortho-substituted-OCH2-fragments in Nitro-Benzo-9-Crown-3 induces low frequency shifts, positive mnDCj, in all 13C NMR resonances which is an indication of the increased shielding in this crown ether. The magnitude of these shifts vary from 15DC7=716 to 54D<...

The application of metabolic precursors for selective stable isotope labeling of aromatic residues in cell-based protein overexpression has already resulted in numerous NMR probes to study the structural and dynamic characteristics of proteins. With anthranilic acid, we present the structurally simplest precursor for exclusive tryptophan side chain labeling. A synthetic route to 13C, 2H isotopo...

Hydrogen isotope ratios of plant lipids are used for paleoclimate reconstruction, but influenced by both source water and biosynthetic processes. Measuring 2H : 1H multiple compounds produced different pathways could allow these effects to be separated, hydrogen fractionations during isoprenoid biosynthesis remain poorly constrained. To investigate how fractionation is molecular exchange betwee...

Intrinsic deuterium-induced isotope effects have been measured and analyzed for compounds having one or two phenyl groups specifically labelled at different positions in a molecule. Some typical examples of directional and orientational dependence of the effects have been given. Different magnitudes of isotope effects that are transmitted in opposite directions have been observed. Thus, in p-2H...

The persistent organic pollutant DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) is still indispensable in the fight against malaria, although DDT and related compounds pose toxicological hazards. Technical DDT contains the dichloro congener DDD (1-chloro-4-[2,2-dichloro-1-(4-chlorophenyl)ethyl]benzene) as by-product, but DDD is also formed by reductive degradation of DDT in the environment...

Mass spectrometry has served as a major tool for the discipline of proteomics to catalogue proteins in an unprecedented scale. With chemical and metabolic techniques for stable isotope labeling developed over the past decade, it is now routinely used as a method for relative quantification to provide valuable information on alteration of protein abundance in a proteome-wide scale. More recently...