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Gene mapping within the T/t complex of the mouse. II. Anomalous position of the H-2 complex in t haplotypes.

Naturally occurring t haplotypes are chromosome 17 polymorphisms that suppress genetic recombination in t/+ heterozygotes over a long distance that includes the H-2 complex. There is strong linkage disequilibrium between t haplotypes and H-2 haplotypes; over 20 independently isolated t chromosomes representing eight different complementation groups share only four H-2 haplotypes. Thus t haplotypes and their associated H-2 loci are inherited en bloc as a "supergene" complex, whose frequency is driven in wild mouse populations by their high transmission from male t heterozygotes. This phenomenon must therefore serve as an important regulator of H-2 polymorphisms. Genes within the region of recombination suppression in t haplotypes have been mapped by crossing-over that occurs readily between two different t haplotypes situated in trans, and by this means we show here that the H-2 complex occupies an anomalous position in t haplotypes, mapping proximal to the locus of tf closely flanked by t-lethal mutations.

Animals↗

Complexation studies on inositol-phosphates. II. Alkali-metal complexes of D-myo-inositol 1,2,6 trisphosphate.

The complexation properties of the D-myo-inositol 1,2,6 trisphosphate (Ins(1,2,6)P3) towards Li+, Na+, K+, Rb+, and Cs+ cations were studied at 25 degrees C in a 0.1 M tetra-n-butylammonium bromide medium. For all cations, mononuclear and protonated species were found. For smaller cations (Li+, Na+, and K+) a dinuclear complex was also put into evidence. The main characteristic of the complexes is its high stability; and of the ligand, its nonselectivity. The Ins(1,2,6)P3-K system was ascertained using Sammartano's method which additionally enabled the influence of various K+ concentrations on the protonations constants to be considered.

Cations, Monovalent↗

An analysis of factors influencing complex water maze learning in rats: effects of task complexity, path order and escape assistance on performance following prenatal exposure to phenytoin.

Three hypotheses on factors determining performance in a complex water maze were tested in rats prenatally exposed to phenytoin. The hypotheses were: 1) that increasing maze complexity would better differentiate experimental effects; in particular, that an expanded version of a maze originally described by Biel would better differentiate groups than Biel's original design; 2) that path order is an important factor determining performance; specifically, that path sequence AB would better differentiate experiments from controls than the opposite order (sequence BA); and 3) that repeated trial failures interfere with learning, a problem putatively prevented by employing assisted (i.e., guided) escape. The specific prediction was that rats tested with assisted escape would learn faster and produce better group differentiation than rats tested with unassisted escape. Pregnant female Sprague-Dawley CD rats were gavaged on days 7-18 of gestation with propylene glycol alone (Control) or containing 100 or 200 mg/kg of phenytoin. Straight channel swimming trials followed by maze trials were begun on separate male/female offspring pairs from each litter on postnatal days 50, 70, or 90. The results confirmed hypothesis 1, i.e., the more complex maze better differentiated phenytoin-related group differences. This was true regardless of whether the phenytoin rats exhibiting circling were included in the analyses or not. The results disconfirmed hypothesis 2, i.e., that path order AB would better differentiate the groups than path order BA. Rather, the data supported the alternate hypothesis, that path order was not a significant determinant of prenatal drug-related maze deficits. This was unchanged regardless of whether phenytoin offspring exhibiting circling were or were not included in the analyses. The implication is that path B alone was sufficient to detect phenytoin's effects on maze performance. Finally, the overall results disconfirmed hypothesis 3, i.e., assisted escape failed to differentiate groups any better than unassisted escape regardless of whether circlers were or were not included in the analyses. However, when more detailed analyses were performed only on rats that failed to find the goal on the first or second trials of path B, an assistance effect was noted. The effect (fewer errors) only occurred in the controls and only on the first two trials of path B. It was concluded that assisted escape was, at best, a minor contribution to improved maze performance.

Animals↗

The 188Re(III)-EDTA complex--a multipurpose starting material for the preparation of relevant 188Re complexes under mild conditions.

An easy and gentle method for the preparation of 188Re(V) complexes with bidentate and tetradentate ligands is described starting from the precursor complex 188Re(III)-EDTA. That complex is prepared at room temperature in acidic solution and reacts by a combined re-oxidation/ligand exchange reaction with appropriate ligands like DMSA or ECD (DMSA = dimercapto succinic acid, ECD = L,L-ethylene dicysteine diethyl ester) or en, tau, and cyclam (en = ethylene diamine, tau = 1,4,8,11-tetraazaundecane, cyclam = 1,4,8,11-tetraazacyclo-tetradecane) to the 188Re(V)-oxo- and dioxocomplexes, respectively. The chelates were unambiguously identified by chromatographic comparison with spectroscopically characterised samples or known 99mTc-kit reconstitutions. The reaction succeeds under mild conditions (room temperature, short time, neutral or weak basic solutions) with high yields and has potential for labelling of sensitive biomolecules with 188Re.

Journal Article↗

Anc1 interacts with the catalytic subunits of the general transcription factors TFIID and TFIIF, the chromatin remodeling complexes RSC and INO80, and the histone acetyltransferase complex NuA3.

The Anc1 protein co-purifies with general transcription factors, chromatin remodeling complexes, and histone modification enzymes and is required for efficient transcription in yeast. We show here that Anc1 interacts with Tsm1, Tfg1, Sth1, Ino80, and Sas3 that are, respectively, the catalytic subunits of the general transcription factors TFIID and TFIIF, of the chromatin remodeling complexes RSC and INO80, and of the histone H3-acetyltransferase complex NuA3. We show that Anc1 is required for growth on galactose as the sole carbon source, and that it is recruited to the UAS of the GAL1 gene after induction.

Acetyltransferases↗

Expression of the skeletal muscle dystrophin-dystroglycan complex and syntrophin-nitric oxide synthase complex is severely affected in the type 2 diabetic Goto-Kakizaki rat.

The inability of insulin to stimulate glucose metabolism in skeletal muscle fibres is a classic characteristic of type 2 diabetes. Using the non-obese Goto-Kakizaki rat as an established animal model of this type of diabetes, sucrose gradient centrifugation studies were performed and confirmed the abnormal subcellular location of the glucose transporter GLUT4. In addition, this analysis revealed an unexpected drastic reduction in the surface membrane marker beta-dystroglycan, a dystrophin-associated glycoprotein. Based on this finding, a comprehensive immunoblotting survey was conducted which showed a dramatic decrease in the Dp427 isoform of dystrophin and the alpha/beta-dystroglycan subcomplex, but not in laminin, sarcoglycans, dystrobrevin, and excitation-contraction-relaxation cycle elements. Thus, the backbone of the trans-sarcolemmal linkage between the extracellular matrix and the actin membrane cytoskeleton might be structurally impaired in diabetic fibres. Immunohistochemical studies revealed that the reduction in the dystrophin-dystroglycan complex does not induce obvious signs of muscle pathology, and is neither universal in all fibres, nor fibre-type specific. Most importantly, the expression of alpha-syntrophin and the syntrophin-associated neuronal isoform of nitric oxide synthase, nNOS, was demonstrated to be severely reduced in diabetic fibres. The loss of the dystrophin-dystroglycan complex and the syntrophin-nNOS complex in selected fibres suggests a weakening of the sarcolemma, abnormal signalling and probably a decreased cytoprotective mechanism in diabetes. Impaired anchoring of the cortical actin cytoskeleton via dystrophin might interfere with the proper recruitment of the glucose transporter to the surface membrane, following stimulation by insulin or muscle contraction. This may, at least partially, be responsible for the insulin resistance in diabetic skeletal muscles.

Animals↗

Comparison of the shape parameters of DNA-cationic lipid complexes and model polyelectrolyte-lipid complexes.

In this study, we used a rheological method to study the shape of DNA-cationic lipid complexes and model polyelectrolyte-lipid complexes. We introduced two kinds of anionic polyelectrolytes, sodium polygalacturonate (PGU) and sodium dextran sulfate (DSS), of varying size, as models for DNA. The prepared complexes were incubated under laminar flow conditions. The results show the same quantitative relation between the shape parameter of lipoplexes and the length of anionic polyelectrolytes, including DNA. The rheological behavior of PGU and DSS were similar to that of DNA.

Anions↗

Characterization and crystal structure of cadmium(II) halide complexes with amino acids and their derivatives VI. The comparison of crystal structures of cadmium(II) halide complexes with three kinds of piperidine carboxylic acids.

Six cadmium(II) halide complexes with dl-piperidine-2-carboxylic acid (DL-Hpipe-2), dl-piperidine-3-carboxylic acid (DL-Hpipe-3), and piperidine-4-carboxylic acid (Hpipe-4), have been prepared and characterized by means of IR and Raman spectra and thermal analysis. The crystal structures of [CdCl2(DL-Hpipe-2)(H2O)], [CdBr2(DL-Hpipe-3)], and [CdCl2(Hpipe-4)] have been determined by X-ray diffraction. These three complexes have one-dimensional polymer structures bridged by halide atoms. The crystal of [CdCl2(DL-Hpipe-2)(H2O)] is orthorhombic with the space group Pca2(1). The cadmium atom is in an octahedral geometry, ligated by a carboxyl oxygen atom, two bridging chlorine atoms, a terminal chlorine atom, a water molecule and a carboxyl oxygen atom of a neighboring molecule. The carboxyl oxygen atoms of DL-Hpipe-2 are coordinated to two cadmium atoms. The unit cell consists of two types of one-dimensional polymer structures: [CdCl2(D-Hpipe-2)(H2O)] and [CdCl2(L-Hpipe-2)(H2O)]. Therefore, it is better to write [CdCl2(DL-Hpipe-2)(H2O)] as [CdCl2(D-Hpipe-2)(H2O)][CdCl2(L-Hpipe-2)(H2O)]. The crystal structure of [CdBr2(DL-Hpipe-3)] is monoclinic with space group P2(1). The cadmium atom is in a distorted octahedral geometry ligated by two carboxyl oxygen atoms and four bridging bromine atoms. This complex consists of either D-Hpipe-3 or L-Hpipe-3. Therefore [CdBr2(DL-Hpipe-3)] is written as [CdBr2(D or L-Hpipe-3)]. The crystal of [CdCl2(Hpipe-4)] is monoclinic with space group P2(1)/n. The structure is similar to that of [CdBr2(D or L-Hpipe-3)].

Amino Acids↗

Self-assembly and cytotoxicity study of waterwheel-like dinuclear metal complexes: the first metal complexes appended with multiple free hydroxamic acid groups.

Two waterwheel-like dinuclear complexes [M(2)(PHA)(4)(H(2)O)(2)] (M = Cu(II) (1), Zn(II) (2); HPHA = phthal-hydroxamic acid) appended with four free hydroxamic acid groups, namely, free hydroxamic acid metal complexes (FHAMCs) have been synthesized and characterized. The crystal structure of complex 1 was determined by single crystal X-ray diffraction, which adopts the paddlewheel motif with four bidentate carboxylate ligands joining two Cu(II) ions. The relative cytotoxicities of compounds 1 and 2 against SMMC-7721 and HO-8910 cell lines are similar and more predominant than HPHA (IC(50): Cu(II)>Zn(II)>>HPHA). The synergic effect of the bound water molecules, multiple free hydroxamic acid groups and dimetal active sites with bridging carboxylate may have significant impacts on their pharmacological activity. As the prototype for a new class of hydroxamic acid derivatives, the self-assembly of FHAMCs presents a promising new strategy in designing multiple hydroxamic acids with remarkable bioactivities.

Antineoplastic Agents↗

A single subunit of a heterotrimeric CCAAT-binding complex carries a nuclear localization signal: piggy back transport of the pre-assembled complex to the nucleus.

An unresolved question concerns the nuclear localization of the heterotrimeric CCAAT-binding complex, which is evolutionarily conserved in eukaryotic organisms including fungi, plants and mammals. All three subunits are necessary for DNA binding. In the filamentous fungus Aspergillus nidulans the corresponding complex was designated AnCF (A.nidulans CCAAT-binding factor). AnCF consists of the HapB, HapC and HapE subunits. Here, by using various green fluorescent protein constructs, a nuclear localization signal sequence (NLS) of the HapB protein was identified, outside of the evolutionarily conserved domain. HapB-EGFP was transported into the nucleus in both DeltahapC and DeltahapE strains, indicating that its NLS interacts with the import machinery independently of the other Hap subunits. In contrast, HapC-EGFP did not enter the nucleus in the absence of HapE or HapB. A similar finding was made for HapE-EGFP, which did not localize to the nucleus in the absence of HapC or HapB. Addition of the HapB-NLS to either HapC or HapE led to nuclear localization of the respective protein fusions, indicating that both HapC and HapE lack a functional NLS. Furthermore, these data strongly suggest that HapC and HapE have first to form a heterodimer and can be transported only as a heterodimer via the HapB protein into the nucleus. Therefore, the HapB subunit is the primary cargo for the import machinery, while HapC and HapE are transported to the nucleus only as a heterodimer and in complex with HapB via a piggy back mechanism. This enables the cell to provide equimolar concentrations of all subunits to the nucleus.

Amino Acid Sequence↗

Conversion of active promoter-RNA polymerase complexes into inactive promoter bound complexes in E. coli by the transcription effector, ppGpp.

Guanosine tetraphosphate (ppGpp) is a signal of nutritional stress that regulates transcription. An RNA polymerase rudder mutant rpoC (Delta 312-315) is found to suppress ppGpp deficiency phenotypes by restoring both negative and positive activities of promoter fusions in vivo, as if ppGpp were present. Measurements of defects in transcription of the PargT tRNA promoter with mutant RNA polymerase reveal that the mutant enzyme quantitatively mimics the presence of added ppGpp. DNaseI footprints and mobility shifts under RNA polymerization conditions reveal that the promoter-specific transcription defect of the mutant enzyme can be ascribed to the presence of inactive dead-end promoter complexes with features similar to those of a stable closed complex. We propose that formation of such inactive complexes represents an alternative explanation of "stringent RNA polymerase" mutant behavior to those currently published, and it represents a newly discovered mode of action of ppGpp.

DNA Footprinting↗

Spectrophotometric studies of molecular complex formation between water-soluble cobalt(II) Schiff base complex and nucleotides in mixed solvent systems.

The formation constants for 1:1 molecular complex formation between water-soluble cobalt(II) tetradentate Schiff base complex, disodium[{bis(4-methoxy-5-sulfo-salicylaldehyde)-4,5-dimethyl-o-phenylenediiminato}cobalt(II)], Na2[Co(SO3-4-meosal-4,5-dmophen)], and nucleotides, adenosine-5'-triphosphate (ATP) and cytidine-5'-triphosphate (CTP), in mixed solvent systems of ethanol and water with different volume fractions of ethanol and water have been determined spectrophotometrically at constant ionic strength (I = 0.2 mol dm(-3) NaClO4) and temperature 278 K. Trends in the values of formation constants according to the volume fractions of ethanol and water in ethanol and water mixed solvent systems, suggest that the trend of molecular complex formation increases with increasing the volume fraction of ethanol in mixed solvent systems.

Cobalt↗

Crystal structures of cyanide complexes of P450cam and the oxygenase domain of inducible nitric oxide synthase-structural models of the short-lived oxygen complexes.

The crystal structure of the ternary cyanide complex of P450cam and camphor was determined to 1.8A resolution and found to be identical with the structure of the active oxygen complex [I. Schlichting et al., 2000, Science 287, 1615]. Notably, cyanide binds in a bent mode and induces the active conformation that is characterized by the presence of two water molecules and a flip of the carbonyl of the conserved Asp251. The structure of the ternary complex of cyanide, L-arginine, and the oxygenase domain of inducible nitric oxide synthase was determined to 2.4A resolution. Cyanide binds essentially linearly, interacts with L-Arg, and induces the binding of a water molecule at the active site. This water is positioned by backbone interactions, located 2.8A from the nitrogen atom of cyanide, and could provide a proton required for O-O bond scission in the hydroxylation reaction of nitric oxide synthase.

Animals↗

Interactions of cis- and trans-platinum(II) complexes with dehydrogenase enzymes in the presence of different mono- and polynucleotides: evidence for a ternary complex.

The inhibition of several dehydrogenase enzymes by cis- and trans-Pt(NH3)2Cl2 have been measured in the presence of baker yeast ribonucleic acid (RNA), calf thymus and salmon sperm deoxyribonuclic acid (DNA) and several mononucleotides (AMP and ATP). The binding constants for the interaction of the platinum complexes to the nucleotides have been calculated and a comparison of those values to the previously calculated platinum complex-enzyme binding constants strongly suggest that platinum compounds are more tightly bound to the enzymes. The binding of the platinum complexes to most of the enzymes was decreased in the presence of any nucleotide, yet it was observed that when using rabbit muscle (M4) lactate dehydrogenase the mononucleotides reduced the binding to a lesser degree while the polynucleotides actually enhanced the platinum-enzyme interaction. The implications of these interactions are discussed.

Adenosine Monophosphate↗

The dimeric complex of cyclomaltoheptaose with 1,14-tetradecanedioic acid. Comparison with related complexes.

The structure of the complex of cyclomaltoheptaose (beta-cyclodextrin, betaCD) with 1,14-tetradecanedioic acid has been determined and refined to a final R=0.0693 based on 9824 observed reflections. Each diacid molecule threads through two betaCD monomers arranged in dimers thus, forming a [3]pseudorotaxane. The end carboxylic groups of adjacent dimers, far apart and fully hydrated, are associated indirectly through water molecules. The positioning of the carboxylic groups with respect to the betaCD dimer and the H-bonds with water molecules are very similar to these of the corresponding complexes of the diacids with 12 and 13 carbon atoms. The bending in the middle of the aliphatic chain is more prominent, compared to that of the corresponding guests with less carbon atoms, thus the end carboxylic groups stay in the same height of the primary faces of the betaCD dimeric complex. As a consequence of the present structure, more close contacts are observed between calculated H-atoms of the guest and O-atoms of the host inside the cavity. This bending is allowed by the width of the betaCD dimer cavity at the secondary interface region.

Bridged-Ring Compounds↗

1H NMR and fluorescence studies of the complexation of DMPG by wheat non-specific lipid transfer protein. Global fold of the complex.

Plant non-specific lipid transfer proteins (LTPs) are proteins which transfer lipids between membranes in vitro and are believed to be involved in the transport of cutin monomers to the cuticle layer in vivo or in the plant defence against phytopathogens. The complexation of DMPG, a diacyl phospholipid, by wheat ns-LTP, a protein extracted from wheat seeds, was followed by 1H NMR and fluorescence spectroscopy. The global fold of the protein was calculated using the DIANA software package from a list of 968 distance constraints. The internal cavity volume, a feature common to all known ns-LTP structures, was estimated to be 750 A3 using the 'CAVITE' program. This model of the complex was obtained by inserting a lipid molecule in the cavity and was energy minimized. The study showed that the protein fold described for the free form was only weakly affected by the insertion of the bulky lipid. Observation of some intermolecular NOEs between the protein and the lipid glycerol moiety revealed that the cavity entrance was located between residues His35 and Arg44. The resulting solution structure was compared to the crystal structure of the maize ns-LTP/palmitate complex.

Arginine↗

Semi-automatic analysis of proteins and protein complexes by automated enzyme immunoassay after separation by high-performance gel-permeation chromatography. Size distribution of C3-IgG complexes.

Enzyme immunoassay (ELISA) was applied in the establishment of an immunological method for the characterization of column effluents. Serum samples, containing aggregated immunoglobulin (IgG), were separated by high-performance gel-permeation chromatography on a TSK column and fractions were collected directly in 96-well ELISA microplates. IgG was determined on anti-IgG-coated plates, followed by development with biotin-labelled anti-IgG and enzyme-labelled avidin. Complement factor C3 was determined on anti-C3-coated plates, developed with biotin-labelled anti-C3 and enzyme-labelled avidin. Complexes between IgG and complement factor C3 were determined on anti-IgG-coated plates, developed with biotin labelled anti-C3 antibody and enzyme-labelled avidin. Complex formation between C3 and IgG after incubation of serum with aggregated IgG was demonstrated. The methodology described is generally applicable to the analysis of biological components and is uniquely useful for the analysis of complexes between different components.

Animals↗

Trace metal-citric acid complexes as inhibitors of calcification and crystal growth. I. Effects of Fe(III), Cr(III) and Al(III) complexes on calcium phosphate crystal growth.

The crystal growth of calcium phosphate has been shown to be inhibited by a low molecular weight Fe(III)-citric acid complex formed when there are high ratios of citrate:iron at neutral pH in aqueous solution. Synergism with citric acid was also observed in systems containing Al(III) or Cr(III) ion, but different unknown complexes seemed to be involved. These results are in qualitative agreement with a preceding study involving calcification of an organic matrix. Since chelating agents like citric acid can compete with the storage proteins for iron, we suggest that the Fe(III)-citrate complexes studied here may be formed in vivo and participate in regulating biologic calcification.

Aluminum↗