Regeneration and modification of cellulose acetate from cigarette waste: Biomedical potential by encapsulation of tetracycline hydrochloride.

Cigarette waste are pervasive litter on Earth, posing a major threat to organisms and ecosystems. However, these waste contain cellulose acetate (CA) and can be recycled, transforming into raw materials for new products. Polymers like CA can be used in biomedical applications as drug carriers and scaffolds for drug release. In this study, cigarette filters waste was collected, recycled and used for fabricating the nanofibrous membrane of cellulose acetate nanofibers (CFCA) through electrospinning technique. Tetracycline hydrochloride (TC) was encapsulated in the nanofibers to prevent bacterial infections. Various analyses were conducted: Scanning Electron Microscope (SEM), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction Analysis (XRD) and Thermogravimetric analysis (TGA). CA and CFCA exhibited high water uptake properties and exhibited similar breaking stress and strain values. Both CA and CFCA effectively acted as stable drug carriers, with sustained in vitro drug release. Antibacterial activity was demonstrated by the drug-loaded CA and CFCA nanofibers against, Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia coli. Based on their cytotoxicity evaluations on mouse fibroblast cells (L929), CA and CFCA fibrous mats demonstrated no cytotoxicity and similar cell viability results. Consequently, the TC-loaded nanofibers made from CA and CFCA exhibited suitable properties for wound healing applications.

Removal of iodine by dry adsorbents in filtered containment venting system after 10 years of Fukushima accident.

Radioactive iodine is a hazardous fission product and a major concern for public health. Special attention is paid to iodine out of 80 fission products because of its short half-life of 8.02 days, high activity, and potential health hazards like its irreversible accumulation in thyroid gland and ability to cause thyroid cancer locally. Radioactive iodine can get released in the form of aerosols (cesium iodide), elemental iodine, and organic iodide after a nuclear accident and can cause off-site and on-site contamination. Filtered containment venting system (FCVS) is a safety system whose main objective is mitigation of severe accidents via controlled venting and removal of different forms of iodine to ensure safety of people and environment. After nuclear accidents like Fukushima, extensive research has been done on the removal of iodine by using dry scrubbers. This review paper presents research status of iodine removal by dry adsorbents especially after 10 years of Fukushima to assess the progress, research gap, and challenges that require more attention. A good adsorbent should be cost-effective; it should have high selective adsorption towards iodine, high thermal and chemical stability, and good loading capacity; and its adsorption should remain unaffected by aging and the presence of inhibitors like CO, NO2, CH3Cl, H2O, and Cl2 and radiation. Research on different dry adsorbents was discussed, and their capability as a potential filter for FCVS was reviewed on the basis of all the above-mentioned features. Metal fiber filters have been widely used for removal of aerosols especially micro- and nanoscale aerosols. For designing a metal fiber filter, optimal size or combination of sizes of fibers, number of layers, and loading capacity of filter should be decided according to feasibility and requirement. Balance between flow resistance and removal efficiency is also very important. Sand bed filters were successful in retention of aerosols, but they showed low trapping of iodine and no trapping of methyl iodide at all. For iodine and methyl iodide removal, many adsorbents like activated carbon, zeolites, metal organic frameworks (MOFs), porous organic frameworks (POPs), silica, aerogels, titanosilicates, etc. have been used. Impregnated activated carbon showed good results but low auto-ignition temperature and decline in adsorption due to aging and inhibitors like NOx made them less suitable. Silver zeolites have been very successful in methyl iodide and iodine removal, but they are expensive and affected by presence of CO. Titanosilicates, macroreticular resins, and chalcogels were also studied and they showed good adsorption capacities, but their thermal stability was low. Other adsorbents like silica, MOFs, aerogels, and POPs also showed promising results for iodine adsorption and good thermal stability, but very limited or no research is available on their performance in severe accident conditions. This review will be very helpful for researchers to understand the merits and demerits of different types of dry adsorbents, the important operating parameters that need optimization for designing an efficient scrubber, margin of research, and foreseeable challenges in removal of different forms of iodine.

BiP inactivation due to loss of the deAMPylation function of FICD causes a motor neuron disease.

PURPOSE: The chaperone protein BiP is the master regulator of the unfolded protein response in the endoplasmic reticulum. BiP chaperone activity is regulated by the post-translational modification AMPylation, exclusively provided by FICD. We investigated whether FICD variants identified in patients with motor neuron disease could interfere with BiP activity regulation. METHODS: Exome sequencing was performed to identify causative pathogenic variants associated with motor neuron diseases. Functional studies were conducted on fibroblasts from patients to explore the molecular mechanism of the disease. RESULTS: We identified biallelic variants in FICD causing a neurodegenerative disease of upper and lower motor neurons. Affected individuals harbor a specific missense variant, Arg374His, positioned in the catalytic motif of the enzyme and important for adenosine triphosphate binding. The mutated residue abolishes intramolecular interaction with the regulatory residue Glu234, essential to inhibit AMPylation and to promote de-AMPylation by FICD. Consequently, fibroblasts from patients with FICD variants have abnormally increased levels of AMPylated and thus inactivated BiP. CONCLUSION: Loss of BiP chaperone activity in patients likely results in a chronic impairment of the protein quality control system in the endoplasmic reticulum. These findings will guide the development of therapeutic strategies for motoneuron and related diseases linked to proteotoxic stress.

Health risks of arsenic buildup in soil and food crops after wastewater irrigation.

Despite considerable research of arsenic (As) level in ground/drinking water of Pakistan, scarce data is available regarding irrigation water contamination by As and associated health risks. The municipal wastewater is routinely applied for soil irrigation in peri-urban agriculture of the country. Since the wastewater composition/contamination and its allied consequences greatly vary in different areas, therefore, it is imperative to check the possible health risks in areas where untreated wastewater is being applied for food crop production. This study analyzed potential health hazards of As-buildup in soil and food plants irrigated with municipal wastewater growing under natural conditions. Sixteen wastewater irrigation locations were selected in District Vehari. From these sites, a total of 16 wastewater samples, 108 soil samples and 65 plant samples were collected for As analysis. Total As contents in wastewater (5.3-63.6 µg/L), soil (1.4-19.6 mg/kg) and plants (0-6.5 mg/kg) greatly varied with sampling location, soil depths and plant type. Based on total As contents in edible tissues, risk assessment parameters, especially cancer risk factor, showed possible health risks (> 0.0001) for wheat crops for children while no risks for other food crops. The use of multiple and diversified food crops is recommended in the study area to minimize the possible risk of As exposure and poisoning. The study also anticipates some future viewpoints considering the on-ground situation of wastewater use, possible exposure of metal(loid)s to human and associated health concerns at local and global scale.

Cadmium Partitioning, Physiological and Oxidative Stress Responses in Marigold (Calendula calypso) Grown on Contaminated Soil: Implications for Phytoremediation.

Marigold (Calendula calypso) is a multipurpose ornamental plant, but its cadmium (Cd) tolerance and phytoremediation potential is unknown. The proposed study was carried out to unravel Cd partitioning, physiological and oxidative stress responses of C. calypso grown under Cd stress. Plants were grown for four months in pots having different soil Cd levels: 0, 25, 50, 75, and 100 mg kg-1 soil. Plant growth, biomass, photosynthetic pigments, leaf water contents, stomatal conductance, and membrane stability index were not decreased at 25 mg kg-1 Cd. At higher levels of Cd stress, activities of antioxidant enzymes (SOD, APX, CAT, POD) increased to mitigate H2O2 and lipid peroxidation. Cadmium uptake in plants increased with increasing soil Cd levels, and roots accumulated a greater portion of Cd, followed by shoots and flowers, respectively. On the basis of Cd accumulation and its tolerance, it was determined that C. calypso can be successfully grown for phytostabilization of Cd contaminated soils.

Assessment of arsenic exposure by drinking well water and associated carcinogenic risk in peri-urban areas of Vehari, Pakistan.

Arsenic (As) is a highly toxic and carcinogenic element. It has received considerable consideration worldwide in recent years due to its highest toxicity to human, and currently, high concentrations observed in the groundwater. Some recent media and research reports also highlighted possible As contamination of groundwater systems in Pakistan. However, there is a scarcity of data about As contents in groundwater in different areas/regions of the country. Consequently, the current study estimated the As concentration in the groundwater used for drinking purpose in 15 peri-urban sites of district Vehari, Pakistan. In total, 127 groundwater samples were collected and examined for As contents in addition to physicochemical characteristics such as temperature, electrical conductivity, pH, total soluble salts, chloride, carbonates, bicarbonates, sodium, potassium, lithium, calcium and barium. Results indicated that the groundwater samples were not fully fit for drinking purposes with several parameters, especially the alarming levels of As (mean As: 46.9 µg/L). It was found that 83% groundwater samples of peri-urban sites in district Vehari have As concentration greater than WHO lower permissible limit (10 µg/L). The risk assessment parameters (mean hazard quotient: 3.9 and mean cancer risk: 0.0018) also showed possible carcinogenic and non-carcinogenic risks associated with ingestion of As-contaminated groundwater at peri-urban sites. Based on the findings, it is anticipated that special monitoring and management of groundwater is necessary in the studied area in order to curtail the health risks associated with the use of As-contaminated drinking water. Moreover, appropriate remediation and removal of As from groundwater is also imperative for the study area before being used for drinking purpose to avoid As exposure and related risks to the local community.

Interplay between HGAL and Grb2 proteins regulates B-cell receptor signaling.

Human germinal center (GC)-associated lymphoma (HGAL) is an adaptor protein expressed in GC B cells. HGAL regulates cell motility and B-cell receptor (BCR) signaling, processes that are central for the successful completion of the GC reaction. Herein, we demonstrate phosphorylation of HGAL by Syk and Lyn kinases at tyrosines Y80, Y86, Y106Y107, Y128, and Y148. The HGAL YEN motif (amino acids 107-109) is similar to the phosphopeptide motif pYXN used as a binding site to the growth factor receptor-bound protein 2 (Grb2). We demonstrate by biochemical and molecular methodologies that HGAL directly interacts with Grb2. Concordantly, microscopy studies demonstrate HGAL-Grb2 colocalization in the membrane central supramolecular activation clusters (cSMAC) following BCR activation. Mutation of the HGAL putative binding site to Grb2 abrogates the interaction between these proteins. Further, this HGAL mutant localizes exclusively in the peripheral SMAC and decreases the rate and intensity of BCR accumulation in the cSMAC. Furthermore, we demonstrate that Grb2, HGAL, and Syk interact in the same complex, but Grb2 does not modulate the effects of HGAL on Syk kinase activity. Overall, the interplay between the HGAL and Grb2 regulates the magnitude of BCR signaling and synapse formation.

Effect of salinity on cadmium tolerance, ionic homeostasis and oxidative stress responses in conocarpus exposed to cadmium stress: Implications for phytoremediation.

Contamination of soil with salinity and Cd negatively affects growth and productivity of plants. The proposed study has been planned to explore the effects of salinity on Cd uptake, tolerance and phytoremediation potential of conocarpus (Conocarpus erectus L.). One-month-old uniform plants of conocarpus were exposed to 0, 8.9, 44.5, 89 and 178 µM Cd alone or in combination with 0, 100 and 200 mM NaCl in Hoagland's nutrient solution. Results revealed that shoot and root biomasses, leaf water content and pigment content decreased more in response to combination of Cd and salinity compared to Cd alone. The Na+ and Cl- concentrations in shoot and root were not affected by Cd alone, but increased in Cd + salinity treatments. The K+ concentration decreased by Cd alone as well as Cd combination with salinity. Plant Cd uptake increased in the presence of salinity but its translocation from root to shoot remained unaffected. Exposure of plants to Cd alone and Cd + salinity caused oxidative stress via overproduction of H2O2 and inducing lipid peroxidation. The activities of antioxidant enzymes such as SOD, CAT, POD and APX increased to mitigate this oxidative stress. It is concluded that the tolerance of conocarpus against Cd stress is decreased in the presence of salinity due to increased uptake of toxic ions and intensification of oxidative stress. Moreover, the Cd uptake behavior of this tree indicates its suitability for phytostabilization of Cd contaminated saline and non-saline soils.

SCO2 mutations cause early-onset axonal Charcot-Marie-Tooth disease associated with cellular copper deficiency.

Recessive mutations in the mitochondrial copper-binding protein SCO2, cytochrome c oxidase (COX) assembly protein, have been reported in several cases with fatal infantile cardioencephalomyopathy with COX deficiency. Significantly expanding the known phenotypic spectrum, we identified compound heterozygous variants in SCO2 in two unrelated patients with axonal polyneuropathy, also known as Charcot-Marie-Tooth disease type 4. Different from previously described cases, our patients developed predominantly motor neuropathy, they survived infancy, and they have not yet developed the cardiomyopathy that causes death in early infancy in reported patients. Both of our patients harbour missense mutations near the conserved copper-binding motif (CXXXC), including the common pathogenic variant E140K and a novel change D135G. In addition, each patient carries a second mutation located at the same loop region, resulting in compound heterozygote changes E140K/P169T and D135G/R171Q. Patient fibroblasts showed reduced levels of SCO2, decreased copper levels and COX deficiency. Given that another Charcot-Marie-Tooth disease gene, ATP7A, is a known copper transporter, our findings further underline the relevance of copper metabolism in Charcot-Marie-Tooth disease.

Phosphorylation of Tyr188 in the WW domain of YAP1 plays an essential role in YAP1-induced cellular transformation.

The Hippo signaling pathway regulates cellular proliferation and survival, thus exerting profound effects on normal cell fate and tumorigenesis. The pivotal effector of this pathway is YAP1, a transcriptional co-activator amplified in mouse and human cancers where it promotes epithelial-to-mesenchymal transition (EMT) and malignant transformation. The Hippo tumor suppressor pathway has been suggested to inhibit the YAP1 function through serine phosphorylation-induced cytoplasmic retention and degradation. Here we report that the tyrosine188 (Y188) site of YAP1 isoform with 2 WW domains (known as YAP1-2) plays an important role in YAP1-induced cellular transformation. IP-Mass Spectrometry analysis of YAP1 identified the phosphorylation of Y188 but not other tyrosine residues. In contrast to the aberrant 3D acinus formation observed in YAP1-WT transduced cells, overexpression of YAP1-Y188F (non-phosphorylated mimic) displayed normal 3D structures. In addition, knockdown of the endogenous YAP1 in MDA-MB231 breast cancer cells inhibited cell proliferation and migration, which were then successfully rescued by the exogenous YAP1-WT and YAP1-Y188E but not Y188F. Mechanistically, we also demonstrated that YAP1-Y188F had a higher affinity to the upstream negative regulator PTPN14 and was extensively localized in the cytoplasm. Since the Y188 is located in the conserved aromatic core of the WW domain of YAP1, our finding has a wide implication for WW domain signaling in general, where Y phosphorylation may act as a common positive regulator of the complex formation via WW domains. In summary, our results indicate that tyrosine 188 plays an important role in the YAP1-induced cellular transformation and its phosphorylation may intriguingly serve as a positive indicator of YAP1 activation.

A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model.

Vaccination with tumor-associated antigens can induce cancer-specific CD8+ T cells. A recent improvement has been the targeting of antigen to dendritic cells (DC) using antibodies that bind DC surface molecules. This study explored the use of multi-trimers of CD40L to target the gp100 melanoma tumor antigen to DC. The spontaneously-multimerizing gene Surfactant Protein D (SPD) was used to fuse gp100 tumor antigen and CD40L, creating the recombinant protein SPD-gp100-CD40L. This "third generation" DC-targeting vaccine was designed to both target antigen to DC and optimally activate dendritic cells by aggregating CD40 trimers on the DC membrane surface. SPD-gp100-CD40L expressed as a 110kDa protein. Analytical light scattering analysis gave elution data corresponding to 4-trimer and multi-trimer SPD-gp100-CD40L oligomers. The protein was biologically active on dendritic cells and induced CD40-mediated NF-κB signaling. DNA vaccination with SPD-gp100-CD40L plasmid, together with plasmids encoding IL-12p70 and GM-CSF, significantly enhanced survival and inhibited tumor growth in a B16-F10 melanoma model. Expression of gp100 and SPD-CD40L as separate molecules did not enhance survival, highlighting the requirement to encode gp100 within SPD-CD40L for optimal vaccine activity. These data support a model where DNA vaccination with SPD-gp100-CD40L targets gp100 to DC in situ, induces activation of these DC, and generates a protective anti-tumor response when given in combination with IL-12p70 and GM-CSF plasmids.

Structural and Functional Diversity of Estrogen Receptor Ligands.

Estrogen receptors, comprised of ERα and ERß isoforms in mammals, act as ligandmodulated transcription factors and orchestrate a plethora of cellular functions from sexual development and reproduction to metabolic homeostasis. Herein, I revisit the structural basis of the binding of ERα to DNA and estradiol in light of the recent discoveries and emerging trends in the field of nuclear receptors. A particular emphasis of this review is on the chemical and structural diversity of an everincreasing repertoire of physiological, environmental and synthetic ligands of estrogen receptors that ultimately modulate their interactions with cognate DNA located within the promoters of estrogenresponsive genes. In particular, modulation of estrogen receptors by small molecule ligands represents an important therapeutic goal toward the treatment of a wide variety of human pathologies including breast cancer, cardiovascular disease, osteoporosis and obesity. Collectively, this article provides an overview of a wide array of small organic and inorganic molecules that can fine-tune the physiological function of estrogen receptors, thereby bearing a direct impact on human health and disease.

Structural insights into the functional versatility of WW domain-containing oxidoreductase tumor suppressor.

Recent work on WW domain-containing oxidoreductase (WWOX) tumor suppressor is beginning to shed new light on both the molecular mechanism of action of its WW domains as well as the contiguous catalytic domain. Herein, the structural basis underlying the ability of WW1 domain to bind to various physiological ligands and how the orphan WW2 tandem partner synergizes its ligand binding in the context of WW1-WW2 tandem module of WWOX is discussed. Notably, the WW domains within the WW1-WW2 tandem module physically associate so as to adopt a fixed spatial orientation relative to each other. In this manner, the association of WW2 domain with WW1 hinders ligand binding to the latter. Consequently, ligand binding to WW1 domain not only results in the displacement of WW2 lid but also disrupts the fixed orientation of WW domains in the liganded conformation. Equally importantly, structure-guided functional approach suggests that the catalytic domain of WWOX likely serves as a retinal oxidoreductase that catalyzes the reversible oxidation and reduction of all-trans-retinal. Collectively, this review provides structural insights into the functional versatility of a key signaling protein with important implications on its biology.

Allostery mediates ligand binding to WWOX tumor suppressor via a conformational switch.

While being devoid of the ability to recognize ligands itself, the WW2 domain is believed to aid ligand binding to the WW1 domain in the context of a WW1-WW2 tandem module of WW domain-containing oxidoreductase (WWOX) tumor suppressor. In an effort to test the generality of this hypothesis, we have undertaken here a detailed biophysical analysis of the binding of WW domains of WWOX alone and in the context of the WW1-WW2 tandem module to an array of putative proline-proline-x-tyrosine (PPXY) ligands. Our data show that while the WW1 domain of WWOX binds to all ligands in a physiologically relevant manner, the WW2 domain does not. Moreover, ligand binding to the WW1 domain in the context of the WW1-WW2 tandem module is two-to-three-fold stronger than when treated alone. We also provide evidence that the WW domains within the WW1-WW2 tandem module physically associate so as to adopt a fixed spatial orientation relative to each other. Of particular note is the observation that the physical association of the WW2 domain with WW1 blocks access to ligands. Consequently, ligand binding to the WW1 domain not only results in the displacement of the WW2 lid but also disrupts the physical association of WW domains in the liganded conformation. Taken together, our study underscores a key role of allosteric communication in the ability of the WW2 orphan domain to chaperone physiological action of the WW1 domain within the context of the WW1-WW2 tandem module of WWOX.

Novel mutations confirm that COL11A2 is responsible for autosomal recessive non-syndromic hearing loss DFNB53.

Hearing loss (HL) is a major public health issue. It is clinically and genetically heterogeneous.The identification of the causal mutation is important for early diagnosis, clinical follow-up, and genetic counseling. HL due to mutations in COL11A2, encoding collagen type XI alpha-2, can be non-syndromic autosomal-dominant or autosomal-recessive, and also syndromic as in Otospondylomegaepiphyseal Dysplasia, Stickler syndrome type III, and Weissenbacher-Zweymuller syndrome. However, thus far only one mutation co-segregating with autosomal recessive non-syndromic hearing loss (ARNSHL) in a single family has been reported. In this study, whole exome sequencing of two consanguineous families with ARNSHL from Tunisia and Turkey revealed two novel causative COL11A2 mutations, c.109G > T (p.Ala37Ser) and c.2662C > A (p.Pro888Thr). The variants identified co-segregated with deafness in both families. All homozygous individuals in those families had early onset profound hearing loss across all frequencies without syndromic findings. The variants are predicted to be damaging the protein function. The p.Pro888Thr mutation affects a -Gly-X-Y- triplet repeat motif. The novel p.Ala37Ser is the first missense mutation located in the NC4 domain of the COL11A2 protein. Structural model suggests that this mutation will likely obliterate, or at least partially compromise, the ability of NC4 domain to interact with its cognate ligands. In conclusion, we confirm that COL11A2 mutations cause ARNSHL and broaden the mutation spectrum that may shed new light on genotype-phenotype correlation for the associated phenotypes and clinical follow-up.

Ligand binding to WW tandem domains of YAP2 transcriptional regulator is under negative cooperativity.

YES-associated protein 2 (YAP2) transcriptional regulator drives a multitude of cellular processes, including the newly discovered Hippo tumor suppressor pathway, by virtue of the ability of its WW domains to bind and recruit PPXY-containing ligands to specific subcellular compartments. Herein, we employ an array of biophysical tools to investigate allosteric communication between the WW tandem domains of YAP2. Our data show that the WW tandem domains of YAP2 negatively cooperate when binding to their cognate ligands. Moreover, the molecular origin of such negative cooperativity lies in an unfavorable entropic contribution to the overall free energy relative to ligand binding to isolated WW domains. Consistent with this notion, the WW tandem domains adopt a fixed spatial orientation such that the WW1 domain curves outwards and stacks onto the binding groove of the WW2 domain, thereby sterically hindering ligand binding to both itself and its tandem partner. Although ligand binding to both WW domains disrupts such interdomain stacking interaction, they reorient themselves and adopt an alternative fixed spatial orientation in the liganded state by virtue of their ability to engage laterally so as to allow their binding grooves to point outwards and away from each other. In short, while the ability of WW tandem domains to aid ligand binding is well documented, our demonstration that they may also be subject to negative binding cooperativity represents a paradigm shift in our understanding of the molecular action of this ubiquitous family of protein modules.

FAM65B is a membrane-associated protein of hair cell stereocilia required for hearing.

In a large consanguineous Turkish kindred with recessive nonsyndromic, prelingual, profound hearing loss, we identified in the gene FAM65B (MIM611410) a splice site mutation (c.102-1G>A) that perfectly cosegregates with the phenotype in the family. The mutation leads to exon skipping and deletion of 52-amino acid residues of a PX membrane localization domain. FAM65B is known to be involved in myotube formation and in regulation of cell adhesion, polarization, and migration. We show that wild-type Fam65b is expressed during embryonic and postnatal development stages in murine cochlea, and that the protein localizes to the plasma membranes of the stereocilia of inner and outer hair cells of the inner ear. The wild-type protein targets the plasma membrane, whereas the mutant protein accumulates in cytoplasmic inclusion bodies and does not reach the membrane. In zebrafish, knockdown of fam65b leads to significant reduction of numbers of saccular hair cells and neuromasts and to hearing loss. We conclude that FAM65B is a plasma membrane-associated protein of hair cell stereocilia that is essential for hearing.

Molecular basis of the binding of YAP transcriptional regulator to the ErbB4 receptor tyrosine kinase.

The newly discovered transactivation function of ErbB4 receptor tyrosine kinase is believed to be mediated by virtue of the ability of its proteolytically-cleaved intracellular domain (ICD) to physically associate with YAP2 transcriptional regulator. In an effort to unearth the molecular basis of YAP2-ErbB4 interaction, we have conducted a detailed biophysical analysis of the binding of WW domains of YAP2 to PPXY motifs located within the ICD of ErbB4. Our data show that the WW1 domain of YAP2 binds to PPXY motifs within the ICD in a differential manner and that this behavior is by and large replicated by the WW2 domain. Remarkably, while both WW domains absolutely require the integrity of the PPXY consensus sequence, non-consensus residues within and flanking this motif do not appear to be critical for binding. In spite of this shared mode of binding, the WW domains of YAP2 display distinct conformational dynamics in complex with PPXY motifs derived from ErbB4. Collectively, our study lends new insights into the molecular basis of a key protein-protein interaction involved in a diverse array of cellular processes.

Molecular determinants of the binding specificity of BH3 ligands to BclXL apoptotic repressor.

B-cell lymphoma extra-large protein (BclXL) serves as an apoptotic repressor by virtue of its ability to recognize and bind to BH3 domains found within a diverse array of proapoptotic regulators. Herein, we investigate the molecular basis of the specificity of the binding of proapoptotic BH3 ligands to BclXL. Our data reveal that while the BH3 ligands harboring the LXXX[A/S]D and [R/Q]XLXXXGD motif bind to BclXL with high affinity in the submicromolar range, those with the LXXXGD motif afford weak interactions. This suggests that the presence of a glycine at the fourth position (G+4)--relative to the N-terminal leucine (L0) within the LXXXGD motif--mitigates binding, unless the LXXXGD motif also contains arginine/glutamine at the -2 position. Of particular note is the observation that the residues at the +4 and -2 positions within the LXXX[A/S]D and [R/Q]XLXXXGD motifs appear to be energetically coupled-replacement of either [A/S]+4 or [R/Q]-2 with other residues has little bearing on the binding affinity of BH3 ligands harboring one of these motifs. Collectively, our study lends new molecular insights into understanding the binding specificity of BH3 ligands to BclXL with important consequences on the design of novel anticancer drugs.

PNPLA6 mutations cause Boucher-Neuhauser and Gordon Holmes syndromes as part of a broad neurodegenerative spectrum.

Boucher-Neuhäuser and Gordon Holmes syndromes are clinical syndromes defined by early-onset ataxia and hypogonadism plus chorioretinal dystrophy (Boucher-Neuhäuser syndrome) or brisk reflexes (Gordon Holmes syndrome). Here we uncover the genetic basis of these two syndromes, demonstrating that both clinically distinct entities are allelic for recessive mutations in the gene PNPLA6. In five of seven Boucher-Neuhäuser syndrome/Gordon Holmes syndrome families, we identified nine rare conserved and damaging mutations by applying whole exome sequencing. Further, by dissecting the complex clinical presentation of Boucher-Neuhäuser syndrome and Gordon Holmes syndrome into its neurological system components, we set out to analyse an additional 538 exomes from families with ataxia (with and without hypogonadism), pure and complex hereditary spastic paraplegia, and Charcot-Marie-Tooth disease type 2. We identified four additional PNPLA6 mutations in spastic ataxia and hereditary spastic paraplegia families, revealing that Boucher-Neuhäuser and Gordon Holmes syndromes in fact represent phenotypic clusters on a spectrum of neurodegenerative diseases caused by mutations in PNPLA6. Structural analysis indicates that the majority of mutations falls in the C-terminal phospholipid esterase domain and likely inhibits the catalytic activity of PNPLA6, which provides the precursor for biosynthesis of the neurotransmitter acetylcholine. Our findings show that PNPLA6 influences a manifold of neuronal systems, from the retina to the cerebellum, upper and lower motor neurons and the neuroendocrine system, with damage of this protein causing an extraordinarily broad continuous spectrum of associated neurodegenerative disease.