Glutathione S-transferase in human organs

Glutathione S-transferase (GSH-T) distribution has been investigated in human tissues. The relative contribution of each species to total enzyme activity of the various tissues has been compared. "Cationic" (pI greater than 7.5) "neutral" (pI 6-6.5) and "anionic" (pI less than 5.4) species of GSH-T were separated by isoelectric focusing. "Cationic" GSH-Ts (ligandin) quantitated by radioimmunoassay were present in all tissues studied. Highest concentrations were in liver, kidney, duodenum, testis and adrenal. "Neutral" and "anionic" GSH-Ts were not present in every tissue or in every specimen of some tissues studied. Marked inter-organ and inter-individual variation in the relative concentration of the 3 GSH-T species may explain individual and organ susceptibility to drugs and toxins and underlines the need for future studies to examine individual enzymes rather than total activity.     

Isolation and characterization of isoprene mutants of Escherichia coli.

Isoprenoid compounds are found in all organisms. In Escherichia coli the isoprene pathway has three distinct branches: the modification of tRNA; the respiratory quinones ubiquinone and menaquinone; and the dolichols, which are long-chain alcohols involved in cell wall biosynthesis. Very little is known about procaryotic isoprene biosynthesis compared with what is known about eucaryote isoprene biosynthesis. This study approached some of the questions about isoprenoid biosynthesis and regulation in procaryotes by isolating and characterizing mutants in E. coli. Mutants were selected by determining their resistance to low levels of aminoglycoside antibiotics, which require an electron transport chain for uptake into bacterial cells. The mutants were characterized with regard to their phenotypes, map positions, enzymatic activities, and total ubiquinone content. In particular, the enzymes studied were isopentenyldiphosphate delta-isomerase (EC 5.3.3.2), farnesyldiphosphate synthetase (EC 2.5.1.1), and higher prenyl transferases.     

Unrestricted recognition of a nonpeptide antigen by CD8+ cytolytic T lymphocytes

CD8+ murine CTL that are specific for an unusual nonpeptide Ag, the heme moiety of hemoglobin, have been derived by in vitro stimulation of spleen cells with hemin. Such CTL demonstrate a requirement for the expression of class I Ag on target cells, yet appear to be unrestricted to the extent that both syngeneic and allogeneic targets precoated with hemin are sensitive to lysis. A series of CTL clones with specificity for hemin was derived from C57BL/6 mice. They exhibited the same type of promiscuous recognition that was observed in CTL populations from a number of different strains. The possibility that hemin acts as a nonspecific mediator of lysis by CTL was ruled out by the fact that a variety of CTL populations and clones specific for different Ag did not exhibit hemin-specific lysis. Some explanations offered to explain these results include 1) the possibility that hemin is recognized by binding to a site on the MHC other than the Ag-binding groove, and 2) the possibility that TCR recognition of a rigid molecule, such as hemin, may be less sensitive to polymorphic variation in the MHC than is recognition of a conventional peptide Ag whose conformation may differ significantly when bound to MHC molecules whose sequences differ within the Ag-binding groove.