DnaJ is a molecular chaperone, which not only binds to its various protein substrates, but can also activate the DnaK cochaperone to bind to its various protein substrates as well. DnaJ is a modular protein, which contains a putative zinc finger motif of unknown function. Quantitation of the released Zn(II) ions, upon challenge with p-hydroxymercuriphenylsulfonic acid, and by atomic absorption showed that two Zn(II) ions interact with each monomer of DnaJ. Following the release of Zn(II) ions, the free cysteine residues probably form disulfide bridge(s), which contribute to overcoming the destabilizing effect of losing Zn(II). Supporting this view, infrared and circular dichroism studies show that the DnaJ secondary structure is largely unaffected by the release of Zn(II). Moreover, infrared spectra recorded at different temperatures, as well as scanning calorimetry, show that the Zn(II) ions help to stabilize DnaJ's tertiary structure. An internal 57-amino acid deletion of the cysteine-reach region did not noticeably affect the affinity of this mutant protein, DnaJDelta144-200, to bind DnaK nor its ability to stimulate DnaK's ATPase activity. However, the DnaJDelta144-200 was unable to induce DnaK to a conformation required for the stabilization of the DnaK-substrate complex. Additionally, the DnaJDelta144-200 mutant protein alone was unimpaired in its ability to interact with its final sigma32 transcription factor substrate, but exhibited reduced affinity toward its P1 RepA and lambdaP substrates. Finally, these in vitro results correlate well with the in vivo observed partial inhibition of bacteriophage lambda growth in a DnaJDelta144-200 mutant background.
The effects of temperature and SDS on the three-dimensional organization and secondary structure of beta-glycosidase from the thermophilic archaeon Sulfolobus solfataricus were investigated by CD, IR spectroscopy and differential scanning calorimetry. CD spectra in the near UV region showed that the detergent caused a remarkable change in the protein tertiary structure, and far-UV CD analysis revealed only a slight effect on secondary structure. Infrared spectroscopy showed that low concentrations of the detergent (up to 0.02%) induced slight changes in the enzyme secondary structure, whereas high concentrations caused the alpha-helix content to increase at high temperatures and prevented protein aggregation.
The scallop Adamussium colbecki can be profitably used for monitoring Antarctic coastal environments but its utility
would be increased if chemical analyses of pollutants were integrated with data on their biological effects. Since oxidative stress is
a common pathway of toxicity induced by xenobiotics, a preliminary biochemical characterization was carried out on the
antioxidant system of this species and baseline data collected for future assessment of the anthropogenic impact in this remote area.
The digestive gland and gills were investigated for levels of glutathione and the activity of several glutathione-dependent and
antioxidant enzymes: gluthathione reductase, EC 1.6.4.2; glyoxalase I, EC 4.4.1.5; glyoxalase II, EC 3.1.2.6; gluthathione
S-transferases, EC 2.5.1.18; Se-dependent, EC 1.11.1.9 and Se-independent, EC 2.5.1.18 glutathione peroxidases; catalase, EC
1.11.1.6; and superoxide dismutase, EC 1.15.1.1. The same enzymatic activities were measured for comparison in the
Mediterranean molluscs Mytilus galloprovincialis and Pecten jacobaeus. Very high levels of glutathione S-transferases were found
in the digestive gland of both species of scallop compared to mussels, suggesting the importance of different feeding behaviour
among these molluscs. However, catalase activity, much higher in Adamussium colbecki than in the Mediterranean molluscs, may
represent a biochemical adaptation to the Antarctic marine environment with high levels of dissolved oxygen. Enzymes from the
Antarctic species appeared to be generally more active at low temperatures but, with a few exceptions, their activities increased at
higher temperatures
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