The role of arginine 47 in the cyclization and coupling reactions of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 - Implications for product inhibition and product specificity

Cyclodextrin glycosyltransferase (CGTase) (EC 2.4.1.19) is used for the industrial production of cyclodextrins. Its application, however, is hampered by the limited cyclodextrin product specificity and the strong inhibitory effect of cyclodextrins on CGTase activity. Recent structural studies have identified Arg47 in the Bacillus circulans strain 251 CGTase as an active-site residue interacting with cyclodextrins, but not with linear oligosaccharides. Arg47 thus may specifically affect CGTase reactions with cyclic substrates or products. Here we show that mutations in Arg47 (to Leu or Gln) indeed have a negative effect on the cyclization and coupling activities; Arg47 specifically stabilizes the oligosaccharide chain in the transition state for these reactions. As a result, the mutant proteins display a shift in product specificity towards formation of larger cyclodextrins. As expected, both mutants also showed lower affinities for cyclodextrins in the coupling reaction, and a reduced competitive (product) inhibition of the disproportionation reaction by cyclodextrins. Both mutants also provide valuable information about the processes taking place during cyclodextrin production assays. Mutant Arg47-->Leu displayed an increased hydrolyzing activity, causing accumulation of increasing amounts of short oligosaccharides in the reaction mixture, which resulted in lower final amounts of cyclodextrins produced from starch. Interestingly, mutant Arg47-->Gln displayed an increased ratio of cyclization/coupling and a decreased hydrolyzing activity. Due to the decreased coupling activity, which especially affects the production of larger cyclodextrins, this CGTase variant produced the various cyclodextrins in a stable ratio in time. This feature is very promising for the industrial application of CGTase enzymes with improved product specificity

Copy number variation in a hospital-based cohort of children with epilepsy

Objective: To evaluate the diagnostic yield of microarray analysis in a hospital-based cohort of children with seizures and to identify novel candidate genes and susceptibility loci for epilepsy. Methods: Of all children who presented with their first seizure in the University Medical Center Groningen (January 2000 through May 2013) (n = 1,368), we included 226 (17%) children who underwent microarray analysis before June 2014. All 226 children had a definite diagnosis of epilepsy. All their copy number variants (CNVs) on chromosomes 1-22 and X that contain protein-coding genes and have a prevalence of <1% in healthy controls were evaluated for their pathogenicity. Results: Children selected for microarray analysis more often had developmental problems (82% vs. 25%, p < 0.001), facial dysmorphisms (49% vs. 8%, p < 0.001), or behavioral problems (41% vs. 13%, p < 0.001) than children who were not selected. We found known clinically relevant CNVs for epilepsy in 24 of the 226 children (11%). Seventeen of these 24 children had been diagnosed with symptomatic focal epilepsy not otherwise specified (71%) and five with West syndrome (21%). Of these 24 children, many had developmental problems (100%), behavioral problems (54%) or facial dysmorphisms (46%). We further identified five novel CNVs comprising four potential candidate genes for epilepsy:MYT1L, UNC5D, SCN4B,andNRXN3. Significance: The 11% yield in our hospital-based cohort underscores the importance of microarray analysis in diagnostic evaluation of children with epilepsy

Potential Dental Biofilm Inhibitors: Dynamic Combinatorial Chemistry Affords Sugar-Based Molecules that Target Bacterial Glucosyltransferase

We applied dynamic combinatorial chemistry (DCC) to find novel ligands of the bacterial virulence factor glucosyltransferase (GTF) 180. GTFs are the major producers of extracellular polysaccharides, which are important factors in the initiation and development of cariogenic dental biofilms. Following a structure-based strategy, we designed a series of 36 glucose- and maltose-based acylhydrazones as substrate mimics. Synthesis of the required mono- and disaccharide-based aldehydes set the stage for DCC experiments. Analysis of the dynamic combinatorial libraries (DCLs) by UPLC-MS revealed major amplification of four compounds in the presence of GTF180. Moreover, we found that derivatives of the glucose-acceptor maltose at the C1-hydroxy group act as glucose-donors and are cleaved by GTF180. The synthesized hits display medium to low binding affinity (KD values of 0.4–10.0 mm) according to surface plasmon resonance. In addition, they were investigated for inhibitory activity in GTF-activity assays. The early-stage DCC study reveals that careful design of DCLs opens up easy access to a broad class of novel compounds that can be developed further as potential inhibitors

Quantum planes and quantum cylinders from Poisson homogeneous spaces

Quantum planes and a new quantum cylinder are obtained as quantization of Poisson homogeneous spaces of two different Poisson structures on classical Euclidean group E(2).Comment: 13 pages, plain Tex, no figure

Quantum symmetric pairs and representations of double affine Hecke algebras of type C∨CnC^\vee C_n

We build representations of the affine and double affine braid groups and Hecke algebras of type $C^\vee C_n$, based upon the theory of quantum symmetric pairs $(U,B)$. In the case $U=U_q(gl_N)$, our constructions provide a quantization of the representations constructed by Etingof, Freund and Ma in arXiv:0801.1530, and also a type $BC$ generalization of the results in arXiv:0805.2766.Comment: Final version, to appear in Selecta Mathematic

Purification and characterization of an NAD+-linked formaldehyde dehydrogenase from the facultative RuMP cycle methylotroph Arthrobacter P1

When Arthrobacter P1 is grown on choline, betaine, dimethylglycine or sarcosine, an NAD+-dependent formaldehyde dehydrogenase is induced. This formaldehyde dehydrogenase has been purified using ammonium sulphate fractionation, anion exchange- and hydrophobic interaction chromatography. The molecular mass of the native enzyme was 115 kDa ± 10 kDa. Gel electrophoresis in the presence of sodium dodecyl sulphate indicated that the molecular mass of the subunit was 56 kDa ± 3 kDa, which is consistent with a dimeric enzyme structure. After ammonium sulphate fractionation the partially purified enzyme required the addition of a reducing reagent in the assay mixture for maximum activity. The enzyme was highly specific for its substrates and the Km values were 0.10 and 0.80 mM for formaldehyde and NAD+, respectively. The enzyme was heat-stable at 50°C for at least 10 min and showed a broad pH optimum of 8.1 to 8.5. The addition of some metal-binding compounds and thiol reagents inhibited the enzyme activity

Патопсихологические особенности и закономерности развития органических психических расстройств при болезни Паркинсона

Проанализированы особенности эмоционально−потребностной сферы, выраженность личностных особенностей, типы отношения к болезни у пациентов с болезнью Паркинсона (БП) и психическими расстройствами. Выявлены патопсихологические факторы формирования органического депрессивного расстройства (F06.36), органического тревожного расстройства (F06.4), органического эмоционально−лабильного расстройства (F06.6), описаны механизмы их патогенеза. Относительно деменции (F02.3) у больных БП единого патопсихологического механизма ее формирования не обнаружено, основная роль в ее патогенезе принадлежит органическому поражению головного мозга.Проаналізовано особливості емоційно−потребової сфери, виразність особистісних особливостей, типи ставлення до хвороби у пацієнтів із хворобою Паркінсона (ХП) та психічними розладами. Виявлено патопсихологічні фактори формування органічного депресивного розладу (F06.36), органічного тривожного розладу (F06.4), органічного емоційно−лабільного розладу (F06.6), описано механізми їх патогенезу. Щодо деменції (F02.3) у хворих на ХП єдиного патопсихологічного механізму її формування не виявлено, основна роль в її патогенезі належить органічному ураженню головного мозку.The peculiarities of emotion−need sphere, degree of personality peculiarities, types of attitude to the disease were analyzed in patients with Parkinson's disease (PD) and mental disorders. Pathopsychological factors of forming organic depressive disorder (F06.36), organic anxiety disorder (F06.4), organic emotional−labile disorder (F06.6) were revealed. The mechanisms of their pathogenesis were described. As for dementia (F02.3), uniform pathopsychological mechanism of its formation was not revealed in patients with PD. Main role in its pathogenesis is played by organic brain lesions

Free q-Schrodinger Equation from Homogeneous Spaces of the 2-dim Euclidean Quantum Group

After a preliminary review of the definition and the general properties of the homogeneous spaces of quantum groups, the quantum hyperboloid qH and the quantum plane qP are determined as homogeneous spaces of Fq(E(2)). The canonical action of Eq(2) is used to define a natural q-analog of the free Schro"dinger equation, that is studied in the momentum and angular momentum bases. In the first case the eigenfunctions are factorized in terms of products of two q-exponentials. In the second case we determine the eigenstates of the unitary representation, which, in the qP case, are given in terms of Hahn-Exton functions. Introducing the universal T-matrix for Eq(2) we prove that the Hahn-Exton as well as Jackson q-Bessel functions are also obtained as matrix elements of T, thus giving the correct extension to quantum groups of well known methods in harmonic analysis.Comment: 19 pages, plain tex, revised version with added materia