Biosorption of heavy metals by Saccharomyces cerevisiae: Effects of nutrient conditions

Studies took place to investigate the effects of different nutrient conditions on the biosorption ability and selectivity of heavy metals by Saccharomyces cerevisiae. After having grown in media supplemented with additional glucose, ammonium, phosphate or cysteine, the yeast was exposed to an equimolar solution of lead, cadmium, zinc and copper. Lead removal from a mixed solution was significantly higher than that of copper, followed by zinc and cadmium. Generally, yeasts from cysteine-rich media showed greatest sorption capacity whereas phosphate addition influenced zinc selectivity. In addition, glucose, fructose and sucrose as carbon sources were examined. Cultures grown in glucose had a better uptake than those cultivated with fructose at an incubation time of 30 min.     

单扫描极谱法快速测定胱氨酸的研究及应用

在pH 9.6的H3BO3-NaOH介质中,胱氨酸于-0.56 V(vs.SCE)处产生一灵敏的吸附还原波。用于胱氨酸的测定,波高与0.8~10μmol/L的胱氨酸有线性关系,检测限为0.4μmol/L。许多氨基酸在较高浓度共存情况下不干扰胱氨酸的测定。应用该法测定了人发及胱氨酸片剂中胱氨酸的含量。     

Trans-Reduction of Cerebral Small Vessel Disease Proteins by Notch-Derived EGF-like Sequences

Cysteine oxidation states of extracellular proteins participate in functional regulation and in disease pathophysiology. In the most common inherited dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), mutations in NOTCH3 that alter extracellular cysteine number have implicated NOTCH3 cysteine states as potential triggers of cerebral vascular smooth muscle cytopathology. In this report, we describe a novel property of the second EGF-like domain of NOTCH3: its capacity to alter the cysteine redox state of the NOTCH3 ectodomain. Synthetic peptides corresponding to this sequence (NOTCH3 N-terminal fragment 2, NTF2) readily reduce NOTCH3 N-terminal ectodomain polypeptides in a dose- and time-dependent fashion. Furthermore, NTF2 preferentially reduces regional domains of NOTCH3 with the highest intensity against EGF-like domains 12–15. This process requires cysteine residues of NTF2 and is also capable of targeting selected extracellular proteins that include TSP2 and CTSH. CADASIL mutations in NOTCH3 increase susceptibility to NTF2-facilitated reduction and to trans-reduction by NOTCH3 produced in cells. Moreover, NTF2 forms complexes with the NOTCH3 ectodomain, and cleaved NOTCH3 co-localizes with the NOTCH3 ectodomain in cerebral arteries of CADASIL patients. The potential for NTF2 to reduce vascular proteins and the enhanced preference for it to trans-reduce mutant NOTCH3 implicate a role for protein trans-reduction in cerebrovascular pathological states such as CADASIL.     

氨基酸对葡萄糖-天冬酰胺模拟体系中丙烯酰胺形成的抑制作用研究

目的 研究4种氨基酸对已建立的葡萄糖-天冬酰胺模拟体系中丙烯酰胺生成的影响。方法 以葡萄糖-天冬酰胺组为对照组,在此基础上分别加入半胱氨酸、甘氨酸、赖氨酸和谷氨酸4种氨基酸为实验组,于140℃反应5～30min,测定反应后模拟体系的褐变程度、色差值、生成丙烯酰胺含量和剩余葡萄糖含量。结果 4种氨基酸中,半胱氨酸和谷氨酸对模拟体系中的褐变度和色差值的抑制作用较好;半胱氨酸和赖氨酸对底物葡萄糖的消耗作用较快;半胱氨酸对丙烯酰胺的抑制效果最好,抑制率为(76.00±0.73)%,其他氨基酸的抑制率分别为甘氨酸(38.39±0.44)%、赖氨酸(51.03±3.09)%和谷氨酸(28.76±2.43)%;进一步研究发现,当添加4.2mmol半胱氨酸时,丙烯酰胺抑制率达(95.32±0.47)%。结论 4种氨基酸相比,半胱氨酸对丙烯酰胺形成的抑制效果最好,且对于降低葡萄糖-天冬酰胺模拟体系的褐变度、色差值、葡萄糖含量有较好的效果。     

Cocaprins, β-Trefoil Fold Inhibitors of Cysteine and Aspartic Proteases from Coprinopsis cinerea

We introduce a new family of fungal protease inhibitors with β-trefoil fold from the mushroom Coprinopsis cinerea, named cocaprins, which inhibit both cysteine and aspartic proteases. Two cocaprin-encoding genes are differentially expressed in fungal tissues. One is highly transcribed in vegetative mycelium and the other in the stipes of mature fruiting bodies. Cocaprins are small proteins (15 kDa) with acidic isoelectric points that form dimers. The three-dimensional structure of cocaprin 1 showed similarity to fungal β-trefoil lectins. Cocaprins inhibit plant C1 family cysteine proteases with K_i in the micromolar range, but do not inhibit the C13 family protease legumain, which distinguishes them from mycocypins. Cocaprins also inhibit the aspartic protease pepsin with K_i in the low micromolar range. Mutagenesis revealed that the β2-β3 loop is involved in the inhibition of cysteine proteases and that the inhibitory reactive sites for aspartic and cysteine proteases are located at different positions on the protein. Their biological function is thought to be the regulation of endogenous proteolytic activities or in defense against fungal antagonists. Cocaprins are the first characterized aspartic protease inhibitors with β-trefoil fold from fungi, and demonstrate the incredible plasticity of loop functionalization in fungal proteins with β-trefoil fold.     

Tracking Protein S‐Fatty Acylation with Proteomics

Recently, Hang and co‐workers developed “acyl‐PEG exchange” (APE), which allows the investigation of protein S‐fatty acylation with mass‐tag labelling and gel electrophoresis, methods that are accessible to any biochemistry laboratory.     

Changes in S-allyl cysteine contents and physicochemical properties of black garlic during heat treatment

This study aimed to investigate the effects of temperature on the black garlic manufacturing process. The moisture content, pH, browning intensity, S-allyl cysteine (SAC) content and antioxidant activity, including DPPH radical scavenging activity and reducing power, were determined. The moisture content of garlic gradually decreased throughout the heating process. The rate of moisture removal was higher at high temperatures compared with low temperatures. The pH also decreased more significantly in garlic heated at high temperatures. The browning intensity increased with increasing temperature. The SAC contents of black garlic were significantly different according to heating temperature; the garlic samples heated at a low temperature had a higher SAC contents. Antioxidant activity, as determined by the DPPH radical scavenging activity and reducing power, increased when the garlic was exposed to higher temperatures.     

Synthesis of DNA-Encoded Disulfide- and Thioether-Cyclized Peptides

DNA-encoded chemical library technologies enable the screening of large combinatorial libraries of chemically and structurally diverse molecules, including short cyclic peptides. A challenge in the combinatorial synthesis of cyclic peptides is the final step, the cyclization of linear peptides that typically suffers from incomplete reactions and large variability between substrates. Several efficient peptide cyclization strategies rely on the modification of thiol groups, such as the formation of disulfide or thioether bonds between cysteines. In this work, we established a strategy and reaction conditions for the efficient chemical synthesis of cyclic peptide–DNA conjugates based on linking the side chains of cysteines. We tested two different thiol-protecting groups and found that tert-butylthio (S-tBu) works best for incorporating a pair of cysteines, and we show that the DNA-linked peptides can be efficiently cyclized through disulfide and thioether bond formation. In combination with established procedures for DNA encoding, the strategy for incorporation of cysteines may be readily applied for the generation and screening of disulfide- and thioether-cyclized peptide libraries.