The Acidic Tail of the Cdc34 Ubiquitin-conjugating Enzyme Functions in Both Binding to and Catalysis with Ubiquitin Ligase SCFC^(dc4*)

Ubiquitin ligases, together with their cognate ubiquitin-conjugating enzymes, are responsible for the ubiquitylation of proteins, a process that regulates a myriad of eukaryotic cellular functions. The first cullin-RING ligase discovered, yeast SCF^(Cdc4), functions with the conjugating enzyme Cdc34 to regulate the cell cycle. Cdc34 orthologs are notable for their highly acidic C-terminal extension. Here we confirm that the Cdc34 acidic C-terminal tail has a role in Cdc34 binding to SCF^(Cdc4) and makes a major contribution to the submicromolar K_m of Cdc34 for SCF^(Cdc4). Moreover, we demonstrate that a key functional property of the tail is its acidity. Our analysis also uncovers an unexpected new function for the acidic tail in promoting catalysis. We demonstrate that SCF is functional when Cdc34 is fused to the C terminus of Cul1 and that this fusion retains partial function even when the acidic tail has been deleted. The Cdc34-SCF fusion proteins that lack the acidic tail must interact in a fundamentally different manner than unfused SCF and wild type Cdc34, demonstrating that distinct mechanisms of E2 recruitment to E3, as is seen in nature, can sustain substrate ubiquitylation. Finally, a search of the yeast proteome uncovered scores of proteins containing highly acidic stretches of amino acids, hinting that electrostatic interactions may be a common mechanism for facilitating protein assembly

A review of acid recovery from acidic mining waste solutions using solvent extraction

This is the peer reviewed version of the following article: Uchenna Kesieme, Andreas Chrysanthou, Maurizio Catulli, and Chu Yong Cheng, ‘A review of acid recovery from acidic mining waste solutions using solvent extraction’, Journal of Chemical Technology and Biotechnology, (2018), which has been published in final form at https://doi.org/10.1002/jctb.5728. Under embargo until 1 July 2019. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.The minerals industry is increasingly being forced by regulatory and cost pressures to reduce the amount of liquid acidic waste they produce. This requires a strong focus on waste reduction by recycling, regeneration and reuse. Four mineral acids were examined for recovery from waste acidic solutions including H 2SO 4, HNO 3, HCl, and H 3PO 4. The selection of the optimal extractant for acid recovery was based on extraction, stripping and scrubbing efficiencies. The extractants suitable for the recovery of H 2SO 4 and HCl are in the order of TEHA > Cyanex 923 > TBP > Alamine 336. TEHA has the highest degree of acid extraction and stripping compared with Cyanex 923 and almost 99% of the acid can be stripped. Alamine 336 can extract higher acid (for H 2SO 4 and HCl systems) than Cyanex 923 and TBP. However loaded acid for Alamine 336 system cannot be stripped using water at 60°C. For the recovery of nitric and phosphoric acids from acidic waste effluents, TBP was the best option. This work clearly demonstrates that extractant suitable for acid extraction may not be suitable for its recovery. However such extractant may be applied for the removal of acid from any waste acidic solution sacrificing the back extraction of the loaded acid. The effective implementation of options for acid recovery was examined to improve sustainability in the mineral industry.Peer reviewe

Interaction of a symmetrical α,α',δ,δ'-Tetramethyl-cucurbit[6]uril with Ln³⁺ : potential applications for isolation of lanthanides

The interaction of a symmetrical α,α′,δ,δ′-tetramethyl-cucurbit[6]uril (TMeQ[6]) with a series of lanthanide cations (Ln³⁺) was investigated in neutral water and in acidic solution. Analysis by single crystal X-ray diffraction revealed that different isomorphous families formed under different synthetic conditions. Such differences in the interaction between TMeQ[6] and Ln³⁺ could potentially be used for isolating heavier Ln³⁺ from their lighter counterparts in neutral solution, and lighter lanthanide cations from their heavier counterparts in acidic solution

A field study of lead phytoextraction by various scented Pelargonium cultivars

Phytoremediation appears to be a promising technique for metal soil clean up, although its successful application on a large scale still remains a challenge. Field experiments for six scented Pelargonium cultivars, conducted on two Pb-contaminated calcareous and acidic soils, revealed vigorous plant growth, with no symptoms of morpho-phytotoxicity in spite of high Pb accumulation levels. Lead contents in the harvestable parts of all plants grown on the acidic and more contaminated soil were significantly higher than those grown on the calcareous soil. Three cultivars (Attar of Roses, Clorinda and Atomic Snowflake) are Pb-hyperaccumulator plants: they accumulated more than 1000 mg Pb kg1 DW, with high biomass produced

Electrostatic Steering Accelerates C3d:CR2 Association.

Electrostatic effects are ubiquitous in protein interactions and are found to be pervasive in the complement system as well. The interaction between complement fragment C3d and complement receptor 2 (CR2) has evolved to become a link between innate and adaptive immunity. Electrostatic interactions have been suggested to be the driving factor for the association of the C3d:CR2 complex. In this study, we investigate the effects of ionic strength and mutagenesis on the association of C3d:CR2 through Brownian dynamics simulations. We demonstrate that the formation of the C3d:CR2 complex is ionic strength-dependent, suggesting the presence of long-range electrostatic steering that accelerates the complex formation. Electrostatic steering occurs through the interaction of an acidic surface patch in C3d and the positively charged CR2 and is supported by the effects of mutations within the acidic patch of C3d that slow or diminish association. Our data are in agreement with previous experimental mutagenesis and binding studies and computational studies. Although the C3d acidic patch may be locally destabilizing because of unfavorable Coulombic interactions of like charges, it contributes to the acceleration of association. Therefore, acceleration of function through electrostatic steering takes precedence to stability. The site of interaction between C3d and CR2 has been the target for delivery of CR2-bound nanoparticle, antibody, and small molecule biomarkers, as well as potential therapeutics. A detailed knowledge of the physicochemical basis of C3d:CR2 association may be necessary to accelerate biomarker and drug discovery efforts

An acidic fibroblast growth factor protein generated by alternate splicing acts like an antagonist.

Polymerase chain reaction amplification of cDNA for acidic fibroblast growth factor in several lines of cultured human cells revealed two forms of mRNA. The novel smaller mRNA lacks the entire second coding exon of the acidic fibroblast growth factor gene, whereas the previously identified mRNA consists of three coding exons. The truncated variant of acidic fibroblast growth factor (aFGF') is only 60 amino acids long with an apparent molecular mass of 6.7 kD on sodium dodecyl sulfate gels in contrast to 18 kD for the full-length acidic fibroblast growth factor. aFGF' elicits only minimal fibroblast proliferation and antagonizes the effects of acidic fibroblast growth factor when added exogenously to or when coexpressed with aFGF in BALB/c/3T3 fibroblasts. Thus, the truncated variant of acidic fibroblast growth factor may provide fibroblasts with a unique mechanism for endogenous regulation of their responses to acidic fibroblast growth factor

Pancreaticogastrostomy: Effect of Partial Gastrectomy on the Pancreatic Stump in Rabbits

To assess the influence of digestive juice on the pancreatic stump when pancreaticogastrostomy was performed after pancreatoduodenectomy, the pancreatic stump was anastomosed to the intact stomach (group I), the stomach after partial gastrectomy (group II), or the jejunum (group III) in rabbits, and the nature of the digestive juice at the anastomotic site as well as the histologic changes of the pancreatic tissue were investigated. The digestive juice was highly acidic in group I, slightly acidic in group II, and almost neutral in group III. Histological examination of the pancreatic stump revealed extensive coagulative necrosis and delayed replacement with granulation tissue in group I, while there was less prominent liquefactive necrosis and early replacement with granulation tissue in groups II and III. Intraperitoneal abscess formation around the anastomotic site and atrophic fibrosis of the pancreas (similar to the changes after pancreatic duct ligation) occurred in 27.8% and 46.2% of group I rabbits, respectively, but no such changes were detected in groups II and III (both P &#60; 0.05). These results indicate that the highly acidic gastric juice had a widespread corrosive effect on the anastomosed pancreatic tissue, and that partial gastrectomy may be necessary to prevent anastomotic leakage and pancreatic duct obstruction after pancreaticogastrostomy.</p

Mono-\u3cem\u3eN\u3c/em\u3e-acyl-2,6-diaminopimelic Acid Derivatives: Analysis by Electromigration and Spectroscopic Methods and Examination of Enzyme Inhibitory Activity

Thirteen mono-N-acyl derivatives of 2,6-diaminopimelic acid (DAP)—new potential inhibitors of the dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE; EC 3.5.1.18)—were analyzed and characterized by infrared (IR) and nuclear magnetic resonance (NMR) spectroscopies and two capillary electromigration methods: capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC). Structural features of DAP derivatives were characterized by IR and NMR spectroscopies, whereas CZE and MEKC were applied to evaluate their purity and to investigate their electromigration properties. Effective electrophoretic mobilities of these compounds were determined by CZE in acidic and alkaline background electrolytes (BGEs) and by MEKC in acidic and alkaline BGEs containing a pseudostationary phase of anionic detergent sodium dodecyl sulfate (SDS) or cationic detergent cetyltrimethylammonium bromide (CTAB). The best separation of DAP derivatives, including diastereomers of some of them, was achieved by MEKC in an acidic BGE (500 mM acetic acid [pH 2.54] and 60 mM SDS). All DAP derivatives were examined for their ability to inhibit catalytic activity of DapE from Haemophilus influenzae (HiDapE) and ArgE from Escherichia coli (EcArgE). None of these DAP derivatives worked as an effective inhibitor of HiDapE, but one derivative—N-fumaryl, Me-ester-DAP—was found to be a moderate inhibitor of EcArgE, thereby providing a promising lead structure for further studies on ArgE inhibitors