Evidence for an essential sulfhydryl group at the substrate binding site of the A-system transporter of Ehrlich cell plasma membranes

Plasma membrane suspensions of Ehrlich ascites cells solubilized with cholic acid were used to study the effects of sulfhydryl reagents on Na(+)-dependent amino acid transport. These suspensions were treated with the sulfhydryl binding agents p-chloromercuribenzenesulfonic acid or N-ethylmaleimide prior to reconstitution for the assay of transport activity. The proteoliposomes formed from dissolved membranes treated with p-chloromercuribenzenesulfonic acid showed no Na(+)-dependent alpha-aminoisobutyric acid transport, while N-ethylmaleimide pretreated membranes retained approximately 90% of the original activity. To avoid interference by the N-ethylmaleimide component, further studies were carried out with membranes pretreated with 200 microM N-ethylmaleimide prior to p-chloromercuribenzenesulfonic acid treatment. A concentration of 25 microM p-chloromercuribenzenesulfonic acid inhibited Na(+)-dependent alpha-aminoisobutyric acid transport by 50%. The degree of inhibition was dramatically reduced in the presence of substrates specific for the A transport system. Using an inhibition index to address the efficacy of inhibition in presence and absence of substrates, it could be shown that an index of 1.0 in presence of p-chloromercuribenzenesulfonic acid was reduced to 0.84 with (methylamino)isobutyric acid alone and 0.05 in the presence of 100 mM Na+ and 5 mM (methylamino)isobutyric acid. Na+ alone offered no protection. The results show that sulfhydryl group(s) on the amino acid carrier may be directly involved in substrate binding and that substrate binding sites are functional in the disaggregated membrane state. Furthermore, Na+ directly affects (methylamino)isobutyrate binding, since the degree of protection by the amino acid analogue against p-chloromercuribenzenesulfonic acid inhibition was influenced by the presence of Na+.(ABSTRACT TRUNCATED AT 250 WORDS)     

Toxicity of toluene and o-xylene toAcinetobacter calcoaceticus in starvation-survival mode

Summary Toxicity of toluene and o-xylene was tested with a deep subsurface isolate during 120 day starvation period. During 120 day starvation period, toxicity appeared to increase with cell age until cells reached the starvation mode. This result may have an significant implication toin situ bioremediation applications since subsurface bacterial populations are apparently in starvation-survival mode.     

A comparison of the movements of two species of gadoid in the vicinity of an underwater reef

Two saithe (35 and 38 cm) and two pollack (43 and 44 cm) were tracked simultaneously for 170 h. During the day, saithe generally patrolled over the whole of an underwater reef as part of a school, making occasional excursions off the reef to another smaller reef 250m distant. At night, saithe movements were largely limited to the reef. Pollack covered less than 50% of the reef during the study period, moving only small distances off the reef. Pollack swam more slowly than saithe during the day, but at the same speed at night.     

Specificity of action of adrenocorticotrophin in vitro on ascorbate transport in rat adrenal glands

The inhibition of ascorbate transport by rat adrenal quarters in response to steroidogenesis in vitro was shown to be highly specific with respect to tissue, substrate and steroidogenic agent. The transport system in vitro is capable of net accumulation of ascorbate. The evidence is consistent with the conclusion that ascorbate ;depletion' in the adrenal gland is due to a specific block by corticoids of the uptake of ascorbate.