The HP1a disordered C terminus and chromo shadow domain cooperate to select target peptide partners

Drosophila melanogaster heterochromatin protein 1a (HP1a) is essential for compacted heterochromatin structure and the associated gene silencing. Its chromo shadow domain (CSD) is well known for binding to peptides that contain a PXVXL motif. Heterochromatin protein 2 (HP2) is a non-histone chromosomal protein that associates with HP1a in the pericentric heterochromatin, telomeres, and the fourth chromosome. Using NMR spectroscopy, fluorescence polarization, and site-directed mutagenesis, we identified an LCVKI motif in HP2 that binds to the HP1a CSD. The binding affinity of the HP2 fragment is approximately two orders of magnitude higher than that of peptides from PIWI (with a PRVKV motif), AF10 (with a PLVVL motif), or CG15356 (with LYPLL and LSIVA motifs). To delineate differential interactions of the HP1a CSD, we characterized its structure, backbone dynamics, and dimerization constant. We found that the dimerization constant is bracketed by the affinities of HP2 and PIWI, which dock to the same HP1a homodimer surface. This suggests that HP2, but not PIWI, interaction can drive the homodimerization of HP1a. Interestingly, the integrity of the disordered C-terminal extension (CTE) of HP1a is essential for discriminatory binding, whereas swapping the PXVXL motifs does not confer specificity. Serine phosphorylation at the peptide binding surface of the CSD is thought to regulate heterochromatin assembly. Glutamic acid substitution at these sites destabilizes HP1a dimers, but improves the interaction with both binding partners. Our studies underscore the importance of CSD dimerization and cooperation with the CTE in forming distinct complexes of HP1a.     

Monomer sequence of partially hydrolyzed poly(4-tert-butoxystyrene) and morphology of diblock copolymers composing this polymer sequence as one block

The molecular characteristics of poly(4-tert-butoxystyrene) (O) upon hydrolysis reaction were investigated. It is known that O can be converted into poly(4-hydroxystyrene) (H) through hydrolysis reaction using strong acid. In this study, a set of poly(4-tert-butoxystyrene-co-4-hydroxystyrene)s (O/H copolymers) having various conversion rates, f_Hs, were prepared. Hydrolysis reaction is found to occur uniformly by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) where the average f_H obtained was consistent with that from ¹H NMR though a certain distribution in the number of hydrolyzed units was conceived. Monomer sequence of O/H copolymers was determined by ¹³C NMR and the changes in triad concentration were obtained by spectra subtraction method. As a result, ¹³C NMR reveals that O and H are statistically distributed. To evaluate the effect of hydrolysis on microphase-separated structure, we observed the morphology of partially hydrolyzed poly(4-tert-butylstyrene-block-4-tert-butoxystyrene) (BO) by transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS). Samples with f_H from 0.21 to 0.67 form both lamellar (major component) and cylindrical (minor component) structures reflecting both the statistical manner of hydrolysis reaction and the possible localized distribution of H sequence.     

Supercooling of water droplets in jet aviation fuel

Ice formation in aircraft fuel systems is an ongoing problem with potentially disastrous consequences. Unfortunately, the icing of fuel systems is poorly understood. It is well known that at temperatures below 0 °C particles of H₂O suspended in fuel can exist as crystalline ice or metastable supercooled water. In this paper we show that micron sized water droplets immersed in Jet A-1 aviation fuel can exist in a metastable supercooled state to around −36 °C. In fact, the majority of droplets in our experiments froze homogeneously showing that the fuel itself did not catalyse ice formation. We suggest that H₂O particles will remain in a supercooled liquid state until they come into contact with a suitable solid surface in an aircraft’s fuel system or the temperature falls below the homogeneous freezing limit.     

Discovery of a Short‐Chain Dehydrogenase from Catharanthus roseus that Produces a New Monoterpene Indole Alkaloid

Plant monoterpene indole alkaloids, a large class of natural products, derive from the biosynthetic intermediate strictosidine aglycone. Strictosidine aglycone, which can exist as a variety of isomers, can be reduced to form numerous different structures. We have discovered a short‐chain alcohol dehydrogenase (SDR) from plant producers of monoterpene indole alkaloids (Catharanthus roseus and Rauvolfia serpentina) that reduce strictosidine aglycone and produce an alkaloid that does not correspond to any previously reported compound. Here we report the structural characterization of this product, which we have named vitrosamine, as well as the crystal structure of the SDR. This discovery highlights the structural versatility of the strictosidine aglycone biosynthetic intermediate and expands the range of enzymatic reactions that SDRs can catalyse. This discovery further highlights how a sequence‐based gene mining discovery approach in plants can reveal cryptic chemistry that would not be uncovered by classical natural product chemistry approaches.     

Cyp1 Inhibition Prevents Doxorubicin-Induced Cardiomyopathy in a Zebrafish Heart-Failure Model

Doxorubicin is a highly effective chemotherapy agent used to treat many common malignancies. However, its use is limited by cardiotoxicity, and cumulative doses exponentially increase the risk of heart failure. To identify novel heart failure treatment targets, a zebrafish model of doxorubicin-induced cardiomyopathy was previously established for small-molecule screening. Using this model, several small molecules that prevent doxorubicin-induced cardiotoxicity both in zebrafish and in mouse models have previously been identified. In this study, exploration of doxorubicin cardiotoxicity is expanded by screening 2271 small molecules from a proprietary, target-annotated tool compound collection. It is found that 120 small molecules can prevent doxorubicin-induced cardiotoxicity, including 7 highly effective compounds. Of these, all seven exhibited inhibitory activity towards cytochrome P450 family 1 (CYP1). These results are consistent with previous findings, in which visnagin, a CYP1 inhibitor, also prevents doxorubicin-induced cardiotoxicity. Importantly, genetic mutation of cyp1a protected zebrafish against doxorubicin-induced cardiotoxicity phenotypes. Together, these results provide strong evidence that CYP1 is an important contributor to doxorubicin-induced cardiotoxicity and highlight the CYP1 pathway as a candidate therapeutic target for clinical cardioprotection.     

Divergent Synthesis of Novel Cylindrocyclophanes that Inhibit Methicillin-Resistant Staphylococcus aureus (MRSA)

The cylindrocyclophanes are a family of macrocyclic natural products reported to exhibit antibacterial activity. Little is known about the structural basis of this activity due to the challenges associated with their synthesis or isolation. We hypothesised that structural modification of the cylindrocyclophane scaffold could streamline their synthesis without significant loss of activity. Herein, we report a divergent synthesis of the cylindrocyclophane core enabling access to symmetrical macrocycles by means of a catalytic, domino cross-metathesis-ring-closing metathesis cascade, followed by late-stage diversification. Phenotypic screening identified several novel inhibitors of methicillin-resistant Staphylococcus aureus. The most potent inhibitor has a unique tetrabrominated [7,7]paracyclophane core with no known counterpart in nature. Together these illustrate the potential of divergent synthesis using catalysis and unbiased screening methods in modern antibacterial discovery.     

Glycol-modified silanes: novel possibilities for the synthesis of hierarchically organized (hybrid) porous materials

The preparation of porous hierarchical architectures that have structural features spanning from the nanometer to micrometer and even larger dimensions and that exhibit certain functionalities is one of the new challenging frontiers in materials chemistry. The sol-gel process is one of the most promising synthesis routes toward such materials because it not only offers the possibility to incorporate organic functions into the porous host but also offers the possibility to deliberately tailor the pore structure. In this Account, the opportunities given by the application of novel diol-modified silanes are discussed for the synthesis of hierarchically organized inorganic and also inorganic-organic porous monoliths.     

Cell-Free DNA Fragments as Biomarkers of Islet β-Cell Death in Obesity and Type 2 Diabetes

Type 2 diabetes (T2D) typically occurs in the setting of obesity and insulin resistance, where hyperglycemia is associated with decreased pancreatic β-cell mass and function. Loss of β-cell mass has variably been attributed to β-cell dedifferentiation and/or death. In recent years, it has been proposed that circulating epigenetically modified DNA fragments arising from β cells might be able to report on the potential occurrence of β-cell death in diabetes. Here, we review published literature of DNA-based β-cell death biomarkers that have been evaluated in human cohorts of islet transplantation, type 1 diabetes, and obesity and type 2 diabetes. In addition, we provide new data on the applicability of one of these biomarkers (cell free unmethylated INS DNA) in adult cohorts across a spectrum from obesity to T2D, in which no significant differences were observed, and compare these findings to those previously published in youth cohorts where differences were observed. Our analysis of the literature and our own data suggest that β-cell death may occur in subsets of individuals with obesity and T2D, however a more sensitive method or refined study designs are needed to provide better alignment of sampling with disease progression events.     

Moving ion fronts in mixed ionic‐electronic conducting polymer films

The aim of this paper is to analyze moving front dynamics of ions and holes in a planar, mixed ionic‐electronic conducting polymer film. As cations invade the film, holes evacuate; thus, an ionic current is converted to an electronic signal. Recent experiments show that the location of the advancing ion front increases as the square‐root of time, a scaling typically associated with diffusive transport, which is surprising given the large driving voltages utilized. Ionic and electronic transport is modeled via the drift‐diffusion equations. A similarity transformation reduces the governing partial differential equations to ordinary differential equations that are solved numerically. The similarity transformation elucidates the origin of the square‐root‐of‐time front scaling. The similarity solution is then compared to the numerical solution of the full drift‐diffusion equations, finding excellent agreement. When compared with experimental data, our model captures the front location; however, qualitative differences between the ion profiles are observed. © 2015 American Institute of Chemical Engineers AIChE J, 61: 1447–1454, 2015     

Structural and Biochemical Studies of Actin in Complex with Synthetic Macrolide Tail Analogues

The actin filament‐binding and filament‐severing activities of the aplyronine, kabiramide, and reidispongiolide families of marine macrolides are located within the hydrophobic tail region of the molecule. Two synthetic tail analogues of aplyronine C (SF‐01 and GC‐04) are shown to bind to G‐actin with dissociation constants of (285±33) and (132±13) nM , respectively. The crystal structures of actin complexes with GC‐04, SF‐01, and kabiramide C reveal a conserved mode of tail binding within the cleft that forms between subdomains (SD) 1 and 3. Our studies support the view that filament severing is brought about by specific binding of the tail region to the SD1/SD3 cleft on the upper protomer, which displaces loop‐D from the lower protomer on the same half‐filament. With previous studies showing that the GC‐04 analogue can sever actin filaments, it is argued that the shorter complex lifetime of tail analogues with F‐actin would make them more effective at severing filaments compared with plasma gelsolin. Structure‐based analyses are used to suggest more reactive or targetable forms of GC‐04 and SF‐01, which may serve to boost the capacity of the serum actin scavenging system, to generate antibody conjugates against tumor cell antigens, and to decrease sputum viscosity in children with cystic fibrosis.