The Importance of Polarity in the Evolution of the K+ Binding Site of Pyruvate Kinase

In a previous phylogenetic study of the family of pyruvate kinase, we found one cluster with Glu117 and another with Lys117. Those sequences with Glu117 have Thr113 and are K⁺-dependent, whereas those with Lys117 have Leu113 and are K⁺-independent. The carbonyl oxygen of Thr113 is one of the residues that coordinate K⁺ in the active site. Even though the side chain of Thr113 does not participate in binding K⁺, the strict co-evolution between position 117 and 113 suggests that T113 may be the result of the evolutionary pressure to maintain the selectivity of pyruvate kinase activity for K⁺. Thus, we explored if the replacement of Thr113 by Leu alters the characteristics of the K⁺ binding site. We found that the polarity of the residue 113 is central in the partition of K⁺ into its site and that the substitution of Thr for Leu changes the ion selectivity for the monovalent cation with minor changes in the binding of the substrates. Therefore, Thr113 is instrumental in the selectivity of pyruvate kinase for K⁺.     

Lack of Charge Interaction in the Ion Binding Site Determines Anion Selectivity in the Sodium Bicarbonate Cotransporter NBCe1

The Na/HCO₃ cotransporter NBCe1 is a member of SLC4A transporters that move HCO₃⁻ across cell membranes and regulate intracellular pH or transepithelial HCO₃ transport. NBCe1 is highly selective to HCO₃⁻ and does not transport other anions; the molecular mechanism of anion selectivity is presently unclear. We previously reported that replacing Asp⁵⁵⁵ with a Glu (D555E) in NBCe1 induces increased selectivity to other anions, including Cl⁻. This finding is unexpected because all SLC4A transporters contain either Asp or Glu at the corresponding position and maintain a high selectivity to HCO₃⁻. In this study, we tested whether the Cl⁻ transport in D555E is mediated by an interaction between residues in the ion binding site. Human NBCe1 and mutant transporters were expressed in Xenopus oocytes, and their ability to transport Cl⁻ was assessed by two-electrode voltage clamp. The results show that the Cl⁻ transport is induced by a charge interaction between Glu⁵⁵⁵ and Lys⁵⁵⁸. The bond length between the two residues is within the distance for a salt bridge, and the ionic strength experiments confirm an interaction. This finding indicates that the HCO₃⁻ selectivity in NBCe1 is established by avoiding a specific charge interaction in the ion binding site, rather than maintaining such an interaction.     

Unravelling the Carbohydrate‐Binding Preferences of the Carbohydrate‐Binding Modules of AMP‐Activated Protein Kinase

The β subunit of adenosine monophosphate (AMP)‐activated protein kinase (AMPK), which exists as two isoforms (β1 and β2) in humans, has a carbohydrate‐binding module (CBM) that interacts with glycogen. Although the β1‐ and β2‐CBMs are structurally similar, with strictly conserved ligand‐contact residues, they show different carbohydrate affinities. β2‐CBM shows the strongest affinity for both branched and unbranched oligosaccharides and it has recently been shown that a Thr insertion into β2‐CBM (Thr101) forms a pocket to accommodate branches. This insertion does not explain why β2‐CBM binds all carbohydrates with stronger affinity. Herein, it is shown that residue 134 (Val for β2 and Thr for β1), which does not come into contact with a carbohydrate, appears to account for the affinity difference. Characterisation by NMR spectroscopy, however, suggests that mutant β2‐Thr101Δ/Val134Thr differs from that of β1‐CBM, and mutant β1‐Thr101ins/Thr134Val differs from that of β2‐CBM. Furthermore, these mutants are less stable to chemical denaturation, relative to that of wild‐type β‐CBMs, which confounds the affinity analyses. To support the importance of Thr101 and Val134, the ancestral CBM has been constructed. This CBM retains Thr101 and Val134, which suggests that the extant β1‐CBM has a modest loss of function in carbohydrate binding. Because the ancestor bound carbohydrate with equal affinity to that of β2‐CBM, it is concluded that residue 134 plays an indirect role in carbohydrate binding.     

茴油臭氧化选择性的研究

研究了无水乙醇等溶剂对茴油臭氧化的影响以及茴油中三种主要双键物茴脑、α-蒎烯和草蒿脑臭氧化的选择性。研究结果表明 ,采用混合溶剂效果较好 ,混合溶剂的质量比为无水乙醇∶环己烷 =1∶ 3。臭氧对茴脑有很好的选择性     

On the Implication of Water on Fragment‐to‐Ligand Growth in Kinase Binding Thermodynamics

A ligand‐binding study is presented focusing on thermodynamics of fragment expansion. The binding of four compounds with increasing molecular weight to protein kinase A (PKA) was analyzed. The ligands display affinities between low‐micromolar to nanomolar potency despite their low molecular weight. Binding free energies were measured by isothermal titration calorimetry, revealing a trend toward more entropic and less enthalpic binding with increase in molecular weight. All protein–ligand complexes were analyzed by crystallography and solution NMR spectroscopy. Crystal structures and solution NMR data are highly consistent, and no major differences in complex dynamics across the series are observed that would explain the differences in the thermodynamic profiles. Instead, the thermodynamic trends result either from differences in the solvation patterns of the conformationally more flexible ligand in aqueous solution prior to protein binding as molecular dynamics simulations suggest, or from local shifts of the water structure in the ligand‐bound state. Our data thus provide evidence that changes in the solvation pattern constitute an important parameter for the understanding of thermodynamic data in protein–ligand complex formation.     

中国粗榧提取物除草选择作用初探

以种子萌发法测定中国粗榧乙醇提取物对2种作物、4种蔬菜、4种牧草、8种杂草共18种供试植物种子发芽后幼根及幼芽生长抑制作用.试验结果表明:10 g/L粗榧提取物对灰绿藜种子幼根生长抑制率大于80%,对7种供试作物、蔬菜或牧草种子幼根生长抑制率均小于50%,表现出一定选择性.以药土法测定粗榧提取物对不同幼苗期野燕麦及小麦生长抑制作用选择性.结果表明:0～5、5～10、10～15 mm三种根长期野燕麦幼根对粗榧提取物敏感性均显著大于同期小麦幼根敏感性,表现出一定选择性;粗榧提取物对5～10 mm根长期野燕麦和小麦幼根及幼芽生长抑制作用显著大于其他2个根长期抑制作用,表明5～10 mm根长期是小麦和野燕麦较敏感时期.0～5 mm/b麦幼芽对粗榧提取物敏感性大于同期野燕麦幼芽敏感性;5～10 mm小麦及野燕麦幼芽对粗榧提取物敏感性无显著差异.     

Selectivity of Competitive Multivalent Interactions at Interfaces

The development of synthetic, low‐molecular‐weight ligand receptor systems for the selective control of biomolecular interactions remains a major challenge. Binding of oligohistidine peptides to chelators containing Ni²⁺‐loaded nitrilotriacetic acid (NTA) moieties is one of the most widely used and best‐characterised recognition systems. Recognition units containing multiple NTA moieties (multivalent chelator headgroups, MCHs) recognise oligohistidines with substantially increased binding affinities. Different multivalencies both at the level of the MCH and at that of the oligohistidine ligand provide a powerful means to vary the affinity of the interaction systematically. Here we have explored the selectivity for the binding of different oligohistidines to immobilised MCH. Using microarrays of mono‐, bis‐, tris‐ and tetrakis‐NTA chelators spotted at different surface densities, we explored the ability of these binders to discriminate fluorescently labelled hexa‐ and decahistidine peptides. When hexa‐ and decahistidine were tested alone, the discrimination of ligands showed little dependence either on the nature or on the density of the chelator. In contrast, coincubation of both peptides decreased the affinity of hexahistidine, increased the affinity of decahistidine, and made the binding of decahistidine highly dependent on MCH density. Kinetic binding assays by dual‐colour total internal reflection fluorescence spectroscopy revealed active exchange of His₆ by His₁₀ and confirmed the high selectivity towards His₁₀. Our results establish the key role of surface multivalency for the selectivity of multivalent interactions at interfaces.     

Biochemical Characterization and Mechanistic Analysis of the Levoglucosan Kinase from Lipomyces starkeyi

Levoglucosan kinase (LGK) catalyzes the simultaneous hydrolysis and phosphorylation of levoglucosan (1,6‐anhydro‐β‐d ‐glucopyranose) in the presence of Mg²⁺–ATP. For the Lipomyces starkeyi LGK, we show here with real‐time in situ NMR spectroscopy at 10 °C and pH 7.0 that the enzymatic reaction proceeds with inversion of anomeric stereochemistry, resulting in the formation of α‐d ‐glucose‐6‐phosphate in a manner reminiscent of an inverting β‐glycoside hydrolase. Kinetic characterization revealed the Mg²⁺ concentration for optimum activity (20–50 mm ), the apparent binding of levoglucosan (K_m=180 mm ) and ATP (K_m=1.0 mm ), as well as the inhibition by ADP (K_i=0.45 mm ) and d ‐glucose‐6‐phosphate (IC₅₀=56 mm ). The enzyme was highly specific for levoglucosan and exhibited weak ATPase activity in the absence of substrate. The equilibrium conversion of levoglucosan and ATP lay far on the product side, and no enzymatic back reaction from d ‐glucose‐6‐phosphate and ADP was observed under a broad range of conditions. 6‐Phospho‐α‐d ‐glucopyranosyl fluoride and 6‐phospho‐1,5‐anhydro‐2‐deoxy‐d ‐arabino‐hex‐1‐enitol (6‐phospho‐d ‐glucal) were synthesized as probes for the enzymatic mechanism but proved inactive with the enzyme in the presence of ADP. The pyranose ring flip ⁴C₁→¹C₄ required for 1,6‐anhydro‐product synthesis from d ‐glucose‐6‐phosphate probably presents a major thermodynamic restriction to the back reaction of the enzyme.     

叔丁醇对异丁烯二聚反应选择性的影响

以强酸性阳离子交换树脂为催化剂,在高压釜内考察了加入叔丁醇(TBA)前后异丁烯(IB)二聚反应工艺条件对二异丁烯(DIB)选择性的影响.结果表明.在加入TBA后.降低了IB二聚速率.但提高了DIB选择性.当反应温度为90℃、原料中IB质量分数为17%～20%、TBA质量分数为0.6%、搅拌速率750r/min、反应3.5h后,IB的转化率约为60%,DIB的选择性可达80%左右。     

On the Selectivity of an Insulator-Based Dielectrophoretic Microdevice

Insulator-based dielectrophoresis (iDEP) is a dielectrophoretic mode where non-uniform electric fields are created employing insulating structures. This article presents an evaluation of the selectivity of an iDEP microdevice by employing DC electric fields with mixtures of polystyrene nano and microparticles. Experimental and mathematical modeling work was carried out in order to assess the selectivity of an iDEP microdevice to immobilize only the large particles from a binary mixture. Negative dielectrophoretic trapping was observed and the effects of particle concentration, concentration ratio, size ratio, and magnitude of the applied electric potential (200 to 600 V) on the microdevice selectivity were studied. The results demonstrated that high selectivity can be obtained with iDEP.     

作用于昆虫神经系统杀虫剂的选择性与安全性

农药的安全性愈来愈受到人们的关注,作用于昆虫神经系统杀虫剂占有非常重要的地位。从哺乳动物与昆虫靶标差异介绍了作用于昆虫神经系统杀虫剂的选择性与安全性。     

K-L分子筛对甲苯芳环氯化选择性的影响

以K-L分子筛为催化剂,催化氯化甲苯制备邻/对氯甲苯,考察催化剂用量、粒径、重复使用次数、及溶剂等工艺条件对氯化选择性的影响。较优条件:氯化温度55℃,反应时间4h,催化剂粒径200目,K-L分子筛=10.0%,1,2-二氯乙烷=40.0%。在此条件下,甲苯转化率为99.8%,对氯甲苯选择性为84.1%,其催化性能优于工业常用的硫铁粉催化剂。     

Selectivity Determinants of RHO GTPase Binding to IQGAPs

IQ motif-containing GTPase-activating proteins (IQGAPs) modulate a wide range of cellular processes by acting as scaffolds and driving protein components into distinct signaling networks. Their functional states have been proposed to be controlled by members of the RHO family of GTPases, among other regulators. In this study, we show that IQGAP1 and IQGAP2 can associate with CDC42 and RAC1-like proteins but not with RIF, RHOD, or RHO-like proteins, including RHOA. This seems to be based on the distribution of charged surface residues, which varies significantly among RHO GTPases despite their high sequence homology. Although effector proteins bind first to the highly flexible switch regions of RHO GTPases, additional contacts outside are required for effector activation. Sequence alignment and structural, mutational, and competitive biochemical analyses revealed that RHO GTPases possess paralog-specific residues outside the two highly conserved switch regions that essentially determine the selectivity of RHO GTPase binding to IQGAPs. Amino acid substitution of these specific residues in RHOA to the corresponding residues in RAC1 resulted in RHOA association with IQGAP1. Thus, electrostatics most likely plays a decisive role in these interactions.     

Study of Catalyst Selectivity in the Oxidation of Phenol

The selectivity of -crystalline zirconium phosphate, amorphous zirconium phenylphosphonate, amorphous zirconium molybdenum phosphate and microporous crystalline aluminium methylphosphonate was evaluated in oxidation reactions of phenol. The different selectivities are correlated to the properties and structure of each compound.     

高选择性合成阿斯巴甜的研究

评述了阿斯巴甜的各种化学和生物合成路线,重点讨论了有关高选择性合成阿斯巴甜的方法,并针对当前研究生产现状提出了在国内进一步的建议。     

Computational Selectivity Assessment of Protease Inhibitors against SARS-CoV-2

The pandemic of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a serious global health threat. Since no specific therapeutics are available, researchers around the world screened compounds to inhibit various molecular targets of SARS-CoV-2 including its main protease (M^pro) essential for viral replication. Due to the high urgency of these discovery efforts, off-target binding, which is one of the major reasons for drug-induced toxicity and safety-related drug attrition, was neglected. Here, we used molecular docking, toxicity profiling, and multiple molecular dynamics (MD) protocols to assess the selectivity of 33 reported non-covalent inhibitors of SARS-CoV-2 M^pro against eight proteases and 16 anti-targets. The panel of proteases included SARS-CoV M^pro, cathepsin G, caspase-3, ubiquitin carboxy-terminal hydrolase L1 (UCHL1), thrombin, factor Xa, chymase, and prostasin. Several of the assessed compounds presented considerable off-target binding towards the panel of proteases, as well as the selected anti-targets. Our results further suggest a high risk of off-target binding to chymase and cathepsin G. Thus, in future discovery projects, experimental selectivity assessment should be directed toward these proteases. A systematic selectivity assessment of SARS-CoV-2 M^pro inhibitors, as we report it, was not previously conducted.     

Selectivity of shear rate on chains in polymer combination reaction

The combination reaction of linear low density polyethylene initiated by Dicumyl peroxide at elevated temperature was investigated under different steady shear rates. Gel‐permeation chromatography was used to determine the molecular weight distributions, and time–molecular weight superposition method was used to analyze the viscosity curves. The results showed that lower (or higher) shear rate increased the reaction possibility of shorter (or longer) chains. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 839–842, 2006     

Cr(VI)‐containing wastewater treatment by means of ion exchange on weak‐ and strong‐base anion exchangers

The aim of this research work was a thermodynamic and kinetic study of the retention of Cr(VI) ions from a K₂CrO₄ solution on macroporous weak‐ and strong‐base anion exchangers, Lewatit M 64 A and Lewatit MP 500 A, respectively. Also, the correlations among the ion‐exchange rate, the retention capacity of Cr(VI), and some process parameters were established. The parameters studied mainly were the concentration of Cr(VI) ions and the type of the counterions coupled with active groups from the anion exchangers. The results led to the conclusion that, for the Lewatit M 64 A resin, there is the following order of the Cr(VI) retention capacity: RCl > R₂SO₄ > ROH, while for the strong‐base anion exchanger, the retention capacity for the Cr(VI) ions is different: ROH > RCl > R₂SO₄. In the Cl^? form, both anion exchangers have the same retention‐capacity values. On the other side, the weak‐base anion exchanger in SO and OH^? forms presents the lowest retention‐capacity values. The process kinetics also presents some differences: for the Lewatit M 64 A resin, the ion‐exchange rate has lower values, especially in the OH^? form. This result is attributed to the increase of the OH^? ion concentration in the solution and its presence hinders the dissociation of the active groups of a weak‐base anion exchanger. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 2093–2098, 2002