The transport/phosphorylation of N,N'-diacetylchitobiose in Escherichia coli. Characterization of phospho-IIB(Chb) and of a potential transition state analogue in the phosphotransfer reaction between the proteins IIA(Chb) AND IIB(Chb).

Enzyme II permeases of the phosphoenolpyruvate:glycose phosphotransferase system comprise one to five separately encoded polypeptides, but most contain similar domains (IIA, IIB, and IIC). The phosphoryl group is transferred from one domain to another, with histidine as the phosphoryl acceptor in IIA and cysteine as the acceptor in certain IIB domains. IIB(Chb) is a phosphocarrier in the uptake/phosphorylation of the chitin disaccharide, (GlcNAc)(2) by Escherichia coli and is unusual because it is separately encoded and soluble. Both the crystal and solution structures of a IIB(Chb) mutant (C10S) have been reported. In the present studies, homogeneous phospho-IIB(Chb) was isolated, and the phosphoryl-Cys linkage was established by (31)P NMR spectroscopy. Rate constants for the hydrolysis of phospho-IIB(Chb) plotted versus pH gave the same shape peak reported for the model compound, butyl thiophosphate, but was shifted about 4 pH units. Evidence is presented for a stable complex between homogeneous Cys10SerIIB(Chb) (which cannot be phosphorylated) and phospho-IIA(Chb), but not with IIA(Chb). The complex (a tetramer (3)) contains equimolar quantities of the two proteins and has been chemically cross-linked. It appears to be an analogue of the transition state complex in the reaction: phospho-IIA(Chb) + IIB(Chb) <--> IIA(Chb) + phospho-IIB(Chb). This is apparently the first report of the isolation of a transition state analogue in a protein-protein phosphotransfer reaction.

Accumulation of cadmium in and its effect on bank vole tissues after chronic exposure.

Cadmium is one of many metals that are not physiologically or biochemically essential to organisms. This element is extremely dangerous as it is easily absorbed and remains in tissues for a long time. Long exposure to high doses of cadmium may cause biochemical and functional changes in some critical organs. In this study, wheat grains contaminated with cadmium chloride were used to test the influence of cadmium on male bank voles (Clethrionomys glareolus). Doses used in the experiment were environmentally realistic: 0.25 microg g-1 (control), 15 microg g-1, and 40 microg g-1 cadmium (dry weight). The animals were given cadmium-contaminated food and clean water ad libitum for 3 and 6 months. After these exposures, the animals were killed and the kidneys, liver, and testes from each vole were collected for analyses. The concentrations of Cd, Cu, Zn, and Fe in the tissues were determined with an atomic absorption spectrophotometer. The formalin-fixed testes, kidneys, and part of the liver were embedded in paraffin and then stained with hematoxylin and eosin. Cadmium accumulation in the tissues was directly proportional to dose. The highest cadmium concentrations were found in the kidneys of animals fed the highest dose of cadmium. Histological examination of the tissues revealed some pathological changes in the structure of kidneys, liver, and testes.