Ser84 of human renin contributes to the biphasic pH dependence of the renin-angiotensinogen reaction

The pH dependence of the reaction of various renins was investigated using sheep angiotensinogen as a substrate. Human renin showed two separate peaks, but rat and mouse Ren1 renins showed one peak with a shoulder. A comparison of the predicted subsite residues of human renin with those of rat and mouse Ren1 renins revealed that Arg82, Ser84, Thr85, Ala229, and Thr312 are unique in the human sequence. We examined the possible importance of these residues in the unique pH profile of the human renin reaction by replacing these residues with the corresponding residues of rat renin. The replacement of Ser84 of human renin with Gly changed the pH dependence of the reaction to one peak, similarly to rat and mouse Ren1 renins. Other mutant human renins kept two separate peaks, similarly to wild-type human renin. These results indicate that Ser84 of human renin contributes to the biphasic pH dependence of the renin-angiotensinogen reaction.     

Tyrosine-83 of human renin contributes to biphasic pH dependence of the renin-angiotensinogen reaction.

The pH dependence of the reaction of human renin with sheep angiotensinogen had a couple of peaks at pH 6.5 and 8.7. The renin activity at pH 8.7 was 1.4 times higher than that at pH 6.5. After the substitutions of Phe, Ala, and His for Tyr83 of human renin, the peak at pH 6.5 could be observed but the peak at pH 8.7 disappeared. At pH 6.5, these substitutions reduced the kcat of human renin to 11.1, 1.31, and 3.49% of that of wild-type renin, respectively, while their Km remained at similar levels to that of the wild type. These results indicate that Tyr83 of human renin contributes to the renin-angiotensinogen reaction at both pHs and it is essential for the catalytic reaction particularly at the basic pH.     

Effects of low urea concentrations on protein-water interactions

Solvent properties of aqueous media (dipolarity/polarizability, hydrogen bond donor acidity, and hydrogen bond acceptor basicity) were measured in the coexisting phases of Dextran–PEG aqueous two-phase systems (ATPSs) containing .5 and 2.0 M urea. The differences between the electrostatic and hydrophobic properties of the phases in the ATPSs were quantified by analysis of partitioning of the homologous series of sodium salts of dinitrophenylated amino acids with aliphatic alkyl side chains. Furthermore, partitioning of eleven different proteins in the ATPSs was studied. The analysis of protein partition behavior in a set of ATPSs with protective osmolytes (sorbitol, sucrose, trehalose, and TMAO) at the concentration of .5 M, in osmolyte-free ATPS, and in ATPSs with .5 or 2.0 M urea in terms of the solvent properties of the phases was performed. The results show unambiguously that even at the urea concentration of .5 M, this denaturant affects partitioning of all proteins (except concanavalin A) through direct urea–protein interactions and via its effect on the solvent properties of the media. The direct urea–protein interactions seem to prevail over the urea effects on the solvent properties of water at the concentration of .5 M urea and appear to be completely dominant at 2.0 M urea concentration.     

尿素和包膜尿素对大棚内有害气体浓度变化的影响

通过室内模拟和塑料大棚试验,研究了普通尿素和矿物改性包膜尿素对土壤pH值及大棚内有害气体浓度变化的影响.结果表明,在室内模拟试验条件下,两种氮肥施用初期均导致土壤pH值上升,并于1周后达到最大值,上升幅度超过50％,随后开始下降,至第5周回到初始水平.大棚内施用两种氮肥均使棚内NH₃、NO₂和O₃浓度增加,其中施用普通尿素处理的NH₃、NO₂日均挥发量均大于矿物改性包膜尿素;施用普通尿素处理使大棚内土壤的NH₃、棚内NO₂和O₃的最高浓度达到42.36、41.95和86.00 μg·m^-3·d^-1,3种气体浓度均达到了有害气体伤害植物的临界阈值;NH₃、NO₂挥发强度受棚温和光照强度的影响,O₃浓度随光照强度变化而改变.     

The reaction of lucerne seedlings to high concentrations of manganese

Summary Concentrations of manganese lower than 100 ppm did not induce detectable differential effects in young seedlings of lucerne. Concentrations of 400 ppm or higher were unnecessarily high for inducing differential responses.Germination was not affected by concentrations of less than 1,000 ppm Mn.Hundred ppm Mn retarded the rate of leaf appearance, reduced the size of the unifoliate leaf, decreased the production of dry matter, and induced toxicity symptoms.There were no clear-cut differences between lines in the effects of manganese concentration on the rate of leaf appearance, so this character is not a reliable criterion of tolerance to manganese toxicity. However, reduction in size of the unifoliate leaf and decrease in dry weight by high concentration of Mn differentiated clearly between seedlings in their reaction to toxicity. Symptom ratings and manganese content of the shoot gave supplementary information.Maternal influences were unimportant in assessing the reaction of lucerne seedlings to manganese toxicity, but seed size may sometimes affect characters used to measure the degree of tolerance or susceptibility.     

Transport of urea at low concentrations in Chlamydomonas reinhardi.

Urea transport into the unicellular green alga Chlamydomonas reinhardi was investigated to further our understanding of controls operating on urea catabolism in this organism. Transport into cells grown with acetate and deprived of ammonia is a saturable process, mediated by at least two systems operating maximally at different external urea concentrations. The lower concentration system, with an apparent Km for urea of 5.1 micron, was the object of detailed study. Transport of urea from a saturating concentration (57 micron) into ammonia- and acetate-grown cells freshly suspended in ammonia-limited medium was not detected. Upon further culturing in the absence of ammonia, derepression occurred with transport ability, first appearing at about 1 h , reaching a maximum at about 2 h, and maintaining this maximum at least 5 h. In contrast to this, CO2-grown cells became derepressed more slowly, and maximum transport ability was not maintained. Addition of ammonia or methylamine (5 mM) during nitrogen deprivation prevented further increases in transport ability and caused loss of previously acquired transport ability. Cycloheximide (10 microng/ml) had a similar effect. Energy uncouplers or dark, anaerobic conditions depressed transport. By these criteria, transport from low urea concentrations is mediated by a process that requires protein synthesis and activation by cellular energy, and the process has a rapid rate of turnover and of deactivation by ammonia.