Gypsum scale formation and inhibition kinetics with implications in membrane system.

Water desalination using membrane technology is one of the main technologies to resolve water pollution and scarcity issues. In the membrane treatment process, mineral scale deposition and fouling is a severe challenge that can lead to filtration efficiency decrease, permeate quality compromise, and even membrane damage. Multiple methods have been developed to resolve this problem, such as scale inhibitor addition, product recovery ratio adjustment, periodic membrane surface flushing. The performance of these methods largely depends on the ability to accurately predict the kinetics of mineral scale deposition and fouling with or without inhibitors. Gypsum is one of the most common and troublesome inorganic mineral scales in membrane systems, however, no mechanistic model is available to accurately predict the induction time of gypsum crystallization and inhibition. In this study, a new gypsum crystallization and inhibition model based on the classical nucleation theory and a Langmuir type adsorption isotherm has been developed. Through this model, it is believed that gypsum nucleation may gradually transit from homogeneous to heterogeneous nucleation when the gypsum saturation index (SI) decreases. Such transition is represented by a gradual decrease of surface tension at smaller SI values. This model assumes that the adsorption of inhibitors onto the gypsum nucleus can increase the nucleus superficial surface tension and prolong the induction time. Using the new model, this study accurately predicted the gypsum crystallization induction times with or without nine commonly used scale inhibitors over wide ranges of temperature (25-90 °C), SI (0.04-0.96), and background NaCl concentration (0-6 mol/L). The fitted affinity constants between scale inhibitors and gypsum show a good correlation with those between the same inhibitors and barite, indicating a similar inhibition mechanism via adsorption. Furthermore, by incorporating this model with the two-phase mineral deposition model our group developed previously, this study accurately predicts the gypsum deposition time on the membrane material surfaces reported in the literature. We believe that the model developed in this study can not only accurately predict the gypsum crystallization induction time with or without scale inhibitors, elucidate the gypsum crystallization and inhibition mechanisms, but also optimize the mineral scale control in the membrane filtration system.

Haploinsufficiency of cohesin protease, Separase, promotes regeneration of hematopoietic stem cells in mice.

Cohesin recently emerged as a new regulator of hematopoiesis and leukemia. In addition to cohesin, whether proteins that regulate cohesin's function have any direct role in hematopoiesis and hematologic diseases have not been fully examined. Separase, encoded by the ESPL1 gene, is an important regulator of cohesin's function. Canonically, protease activity of Separase resolves sister chromatid cohesion by cleaving cohesin subunit-Rad21 at the onset of anaphase. Using a Separase haploinsufficient mouse model, we have uncovered a novel role of Separase in hematopoiesis. We report that partial disruption of Separase distinctly alters the functional characteristics of hematopoietic stem/progenitor cells (HSPCs). Although analyses of peripheral blood and bone marrow of Espl1+/Hyp mice broadly displayed unperturbed hematopoietic parameters during normal hematopoiesis, further probing of the composition of early hematopoietic cells in Espl1+/Hyp bone marrow revealed a mild reduction in the frequencies of the Lin- Sca1+ Kit- (LSK) or LSK CD48+ CD150- multipotent hematopoietic progenitors population without a significant change in either long-term or short-term hematopoietic stem cells (HSCs) subsets at steady state. Surprisingly, however, we found that Separase haploinsufficiency promotes regeneration activity of HSCs in serial in vivo repopulation assays. In vitro colony formation assays also revealed an enhanced serial replating capacity of hematopoietic progenitors isolated from Espl1+/Hyp mice. Microarray analysis of differentially expressed genes showed that Separase haploinsufficiency in HSCs (SP-KSL) leads to enrichment of gene signatures that are upregulated in HSCs compared to committed progenitors and mature cells. Taken together, our findings demonstrate a key role of Separase in promoting hematopoietic regeneration of HSCs.

Mechanism of Ribonuclease III Catalytic Regulation by Serine Phosphorylation.

Ribonuclease III (RNase III) is a conserved, gene-regulatory bacterial endonuclease that cleaves double-helical structures in diverse coding and noncoding RNAs. RNase III is subject to multiple levels of control, reflective of its global regulatory functions. Escherichia coli (Ec) RNase III catalytic activity is known to increase during bacteriophage T7 infection, reflecting the expression of the phage-encoded protein kinase, T7PK. However, the mechanism of catalytic enhancement is unknown. This study shows that Ec-RNase III is phosphorylated on serine in vitro by purified T7PK, and identifies the targets as Ser33 and Ser34 in the N-terminal catalytic domain. Kinetic experiments reveal a 5-fold increase in kcat and a 1.4-fold decrease in Km following phosphorylation, providing a 7.4-fold increase in catalytic efficiency. Phosphorylation does not change the rate of substrate cleavage under single-turnover conditions, indicating that phosphorylation enhances product release, which also is the rate-limiting step in the steady-state. Molecular dynamics simulations provide a mechanism for facilitated product release, in which the Ser33 phosphomonoester forms a salt bridge with the Arg95 guanidinium group, thereby weakening RNase III engagement of product. The simulations also show why glutamic acid substitution at either serine does not confer enhancement, thus underscoring the specific requirement for a phosphomonoester.

Combined computational and experimental analysis of a complex of ribonuclease III and the regulatory macrodomain protein, YmdB.

Ribonuclease III is a conserved bacterial endonuclease that cleaves double-stranded(ds) structures in diverse coding and noncoding RNAs. RNase III is subject to multiple levels of control that in turn confer global post-transcriptional regulation. The Escherichia coli macrodomain protein YmdB directly interacts with RNase III, and an increase in YmdB amount in vivo correlates with a reduction in RNase III activity. Here, a computational-based structural analysis was performed to identify atomic-level features of the YmdB-RNase III interaction. The docking of monomeric E. coli YmdB with a homology model of the E. coli RNase III homodimer yields a complex that exhibits an interaction of the conserved YmdB residue R40 with specific RNase III residues at the subunit interface. Surface Plasmon Resonance (SPR) analysis provided a KD of 61 nM for the complex, corresponding to a binding free energy (ΔG) of -9.9 kcal/mol. YmdB R40 and RNase III D128 were identified by in silico alanine mutagenesis as thermodynamically important interacting partners. Consistent with the prediction, the YmdB R40A mutation causes a 16-fold increase in K(D) (ΔΔG = +1.8 kcal/mol), as measured by SPR, and the D128A mutation in both RNase III subunits (D128A/D128'A) causes an 83-fold increase in KD (ΔΔG = +2.7 kcal/mol). The greater effect of the D128A/D128'A mutation may reflect an altered RNase III secondary structure, as revealed by CD spectroscopy, which also may explain the significant reduction in catalytic activity in vitro. The features of the modeled complex relevant to potential RNase III regulatory mechanisms are discussed.