Expression of sialyl Le, sialyl Le, Le and Le glycotopes in secreted human ovarian cyst glycoproteins

Human blood group A, B, H, Ii, Le^a and Le^b antigens and their determinants expressed on ovarian cyst glycoproteins have been studied for over five decades. However, little is known about sialyl Le^x and sialyl Le^a glycotopes, which play essential roles in normal immunity, inflammation, and cancer cell metastasis. Furthermore, Le^x and Le^y were classified as glycotopes of unknown genes. Identification of these Lewis epitopes was hampered by the lack of specific antibodies. In this study, the occurrence of sialyl Le^x, sialyl Le^a, Le^x and Le^y reactivities in cyst glycoproteins was characterized by enzyme-linked immunosorbent assays. The results indicated that most human ovarian cyst glycoproteins carried Le^x (8/25) and/or Le^y (17/25) glycotopes. The expression (epitopes) of the new genes described in previous reports are Le^x and Le^y glycotopes; the reactivities of sialyl Le^x and sialyl Le^a glycotopes in secreted cyst glycoproteins may be affected by the conditions of purification; the relationship between Le^y and human blood group ABH was confirmed; recognition profiles of sialyl Le^x, sialyl Le^a, Le^x and Le^y present in the carbohydrate chains of water-soluble cyst glycoproteins were illustrated; possible attachments of glycotopes to the internal carbohydrate complex of cyst glycoproteins have been reconstructed; proposed biosynthetic pathways for the formation of sialyl Le^a, sialyl Le^x, Le^x, Le^y, ALe^y and BLe^y determinant structures on Type I and Type II core structures of human ovarian cyst glycoproteins are also included in this study.     

Limnological control of brine shrimp population dynamics and cyst production in the Great Salt Lake, Utah

In the Great Salt Lake of Utah, the brine shrimp Artemia franciscanaKellogg is an important food resource for birds and they produce dormant cysts that are harvested and used extensively in the aquaculture industry. We analyzed the limnological factors controlling Artemia growth and cyst production over 12 months in 1994 and 1995. Laboratory experiments showed that inter-brood intervals were highly dependent on temperature and slightly on food level. At optimal temperatures and nutritious food, juveniles reached reproductive size within 7 d in the laboratory. In winter when temperatures were less than 3 °C, Artemia were absent from the lake, phytoplankton abundance was high (13 Chl a g l^–1), and the dominant grazers were ciliated protozoans. In the spring, cysts hatched when phytoplankton was abundant (15–30 g Chl a l^–1), and the Artemia grew and produced large clutches of ovoviviparous eggs. Estimated naupliar production from these eggs was 80 l^–1 from April to May. Despite the high production of nauplii, Artemia densities declined to 8 l^–1by June and the growing shrimp population grazed down the phytoplankton resource to <1 g Chl a l^–1. With the depleted phytoplankton food resource during the summer, Artemia growth slowed, lipid indices decreased, clutch sizes declined, and females switched primarily to oviparous cyst production. During the summer, there was limited production of ovoviviparous eggs, and limited recruitment of juveniles, probably due to low food. Although oviparous reproduction began in June, more than 90% of the cysts were produced after July when female densities had declined to 1.5 l^–1, but nearly all of them were producing cysts. Estimated cyst production was 650000 m^–2, or 4.54 × 10⁶ kg dry weight for the entire lake. The reported commercial harvest took 21% of the 1994 cyst production. That harvest had little impact on the subsequent year's population, as Artemia densities were ultimately controlled by algal production in the lake.     

Light-path length and population density in photoacclimation of Nannochloropsis sp. (Eustigmatophyceae)

Photoacclimation in the marine eustigmatophyte Nannochlropsis sp., used extensively as a food chaincomponent in aquaculture, was studied both in thelaboratory and outdoors. Cell-chlorophyll andcarotenoids were used as markers to assessphotoacclimation to strong light, as well as todecreasing growth irradiance due to cellproliferation. Focusing on practical aspects involvedin mass cultivation, three different approaches wereused as follows: (a) cultures initially exposed to lowlight (150 mol photon m^-2 s^-1) thentransferred to strong light (1000 to 3000 molphoton m^-2 s^-1); (b) initially low celldensity cultures grown in reactors of differentlight-paths, exposed to strong PFD, in the laboratoryand outdoors; (c) initially low or high cell densitycultures exposed to strong light. As has already beenestablished in many reports, cell-chlorophyllrepresented a sensitive parameter in assessing cellresponse to changes in the intensity of the lightsource as well as to modifications in the light regimeto which the cells were exposed. Cell-chlorophyllconcentration sharply decreased initially upontransferring the culture from low PFD cell^-1 tohigh PFD cell^-1 due to either culture dilution(i.e. decrease in cell density and mutual shading) orto an increase in PFD. After some 7 days ofphotoacclimating to 2000 and 3000 mol photonm^-2 s^-1, chlorophyll a content began to riseat a much faster rate than cell number, which alsoincreased in response to the higher irradiance.Cell-chlorophyll in the culture exposed to 2000mol photon m^-2 s^-1 increased afteracclimation earlier and at a faster rate than in theculture exposed to 3000 mol photon m^-2s^-1, indicating the later irradiance affected astronger stress. The length of the reactor's lightpath exerted a decisive effect on cell response tostrong light through its influence on the light regimein the culture. Upon a sharp increase in PFD,carotenoids in the 1-cm reactor increased in muchhigher rate than chlorophyll, compared with the 3-cmlight path reactors. This marked difference in cellresponse to a shift-up in light was attributed to thevast variations in the light regime associated withdifferences in the length of the light path and areal density. Growth oflow cell density cultures ceased temporarily upontransfer to strong light, in contrast with high celldensity cultures transferred to strong light, whichcontinued growth without a lag.     

Estradiol receptor binding to the epithelium of uterine lumen and glands: region- and time-related changes during preimplantation and periimplantation periods studied by autoradiography

The presence and changes of estradiol nuclear binding and related functions in uterine luminal and glandular epithelium were studied before and after blastocyst implantation using receptor autoradiography with ³H-estradiol-17 in association with ³H-thymidine incorporation and immunocytochemical binding of antibody to estrogen receptor ER-. ³H-estradiol nuclear binding is present but variable during days 1.5–7.5 of pregnancy. Sites of strong nuclear binding of ³H-estradiol exhibit strong immunocytochemical staining with ER- antibody. Qualitative and quantitative evaluation of autoradiograms reveal that there is a general increase of nuclear ³H-estradiol binding during the first 3 days after fertilization in both luminal and glandular epithelium. The binding of estradiol is stronger in glandular epithelium from day 2.5 to day 7.5, paralleled by a rise in ³H-thymidine incorporation on day 2.5. By comparison, in the epithelium of the uterine lumen ³H-estradiol nuclear binding is low, but relatively high in epithelial cells at lateral branching of the lumen where the increase in ³H-estradiol binding corresponds to an increased labeling index with ³H-thymidine. A highly differentiated binding of ³H-estradiol to luminal and glandular epithelium was demonstrated with region- and time-specific changes of related effects on cell proliferation, differentiation, and secretion, probably involving involution and remodeling. The strong ³H-estradiol binding to glandular epithelium suggests that estradiol exerts pronounced effects on glandular activities in the periimplantation period.     

Zinc complexes of a new N3O ligand and their biomimetic reactions

The new N₃O ligand 2-(pyridylmethylamino)-3-(3,5-dimethylpyrazol-1-yl)-propionic acid (L1H) was synthesized and converted to L1Zn-Cl and L1Zn-Br. These complexes are tetrameric in the solid state with bridging carboxylate functions. The reaction of deprotonated L1H with zinc nitrate or zinc perchlorate yielded the aqua complexes [L1Zn-OH₂] X with and , which crystallize as carboxylate-bridged dimers. Their deprotonation produced, in situ, the hydroxide complex L1Zn-OH, which acted as a base toward p-nitrophenol and bis(p-nitrophenyl)phosphoric acid resulting in L1Zn-ONit and L1Zn-OPO(ONit)₂. Tris(p-nitrophenyl)phosphate was cleaved hydrolytically by L1Zn-OH, releasing one p-nitrophenyl group. A kinetic investigation of this cleavage reaction under pseudo-first-order conditions has yielded second-order rate constants k″ of 0.9 s⁻¹ M⁻¹ in 50% aqueous DMSO and 4.0 s⁻¹ M⁻¹ in 75% aqueous DMSO.     

Luminescent metalloreceptors with pendant macrocyclic ionophore: Synthesis, characterization, electrochemistry and ion-binding study

Metalloreceptors containing ruthenium(II) bipyridine unit as fluorophore and pendant macrocyclic units as ionophore have been synthesized and their luminescence and electrochemical properties have been investigated. Ion-binding study of these fluoroionophore with the anions F⁻, Cl⁻, Br⁻, I⁻, , , , , CH₃COO⁻, and and cations Na⁺, K⁺, Mg²⁺, Ca²⁺, Zn²⁺, Ba²⁺, Sr²⁺ Cd²⁺, Hg²⁺, Pb²⁺ and Cu²⁺, monitored by luminescence and ¹H NMR spectral changes, reveal strong interactions of and F⁻ for 2 and 3 and of Cu²⁺ only for 3. Luminescence titrations for 2 and 3 have been carried out to determine binding constants (K_s), and the calculated values are in the range 2.85 × 10² to 4.48 × 10⁴ M⁻¹. The ¹H NMR spectral changes for 2 and 3 with the addition of increasing concentration of F⁻ and exhibit substantial low-field shift of the CONH proton indicating its involvement in complex formation with the anions. The adduct of 2 and 3 have been isolated and characterized by ¹H and ³¹P NMR, mass and IR spectroscopy. The results are discussed in light of selectivity, structures of the anion bound complexes and their luminescence property.     

Mononuclear cobalt(II) carboxylate complexes: Synthesis, molecular structure and selective oxygenation study

Some cobalt carboxylate (both mononuclear as well as binuclear) complexes have been prepared by using hindered hydrotris(3,5-diisopropyl-1-pyrazolyl)borate (Tp^iPr2) as supporting ligand. The reaction of [Tp^iPr2Co(NO₃)] (2) with sodium benzoate resulted in the formation of acetonitrile coordinated complex [Tp^iPr2Co(OBz)(CH₃CN)] (3) whereas the reaction of 2 with sodium fluorobenzoate gave coordinately unsaturated five coordinate complex of the type [Tp^iPr2Co(F-OBz)] (4). The oxidation of compound 4 in the presence of 3,5-diisopropylpyrazole resulted in the formation of a unique compound (5) where only one methine carbon of isopropyl group on pyrazole ring of hydrotris(3,5-diisopropyl-1-pyrazolyl)borate oxidized and coordinated with cobalt center. In compound 5, the binding behavior of fluorobenzoate also changes from bidentate to monodentate and the nonbonded oxygen atom formed intramolecular hydrogen bond with the hydrogen atom of the NH fragment of the coordinated . X-ray crystallography and IR studies confirmed the existence of hydrogen bonding in complex 5. The pyrazolato bridged binuclear cobalt(II) complex (6) was prepared by the reaction of hydrated cobalt(II) nitrate, 3,5-diisopropylpyrazole and sodium nitrobenzoate where, each cobalt is four coordinate. The X-ray structure of 6 showed that the NH fragment of terminally coordinated formed intramolecular hydrogen bonding with nonbonded oxygen atom of monodentately coordinated nitrobenzoate.     

Factors controlling long-term changes in soil pools of exchangeable basic cations and stream acid neutralizing capacity in a northern hardwood forest ecosystem

Understanding the factors regulating the concentrations of basic cations in soils and surface waters is critical if rates of recovery are to be predicted in response to decreases in acidic deposition. Using a dynamic simulation model (PnET-BGC), we evaluated the extent to which atmospheric deposition of strong acids and associated leaching by strong anions, atmospheric deposition of basic cations through changes in emissions of particulate matter, and historical forest cutting have influenced soil pools of exchangeable basic cations and the acid-base status of stream water at the Hubbard Brook Experimental Forest (HBEF) in New Hampshire. Historical deposition of basic cations was reconstructed from regression relationships with particulate matter emissions. Simulation results indicate that the combination of these factors has resulted in changes in the percent soil base saturation, and stream pH and acid neutralizing capacity (ANC) from pre-industrial estimates of 20%, 6.3 and 45 eq L^–1, respectively, to current values of 10%, 5.0 and –5 eq L^–1, respectively. These current values fall within the critical thresholds at which forest vegetation and aquatic biotic are at risk from soil and surface water acidification due to acidic deposition. While the deposition of strong acid anions had the largest impact on the acid-base status of soil and stream water, the reduction in deposition of basic cations associated with reductions in particulate emissions was estimated to have contributed about 27% of the depletion in soil Ca²⁺ exchange pool and 15% of the decreases in stream water concentrations of basic cations. Decline in stream water concentrations of basic cation occurred under both increasing and decreasing exchangeable pools, depending on the process controlling the acid base status of the ecosystem. Model calculations suggest that historical forest cutting has resulted in only slight decreases in soil pools of exchangeable basic cations, and has had a limited effect on stream ANC over the long-term.     

Regulation of adenosine 5′-phosphosulfate sulfotransferase activity by H2S and cyst(e)ine in primary leaves of Phaseolus vulgaris L.

During chloroplast development in the primary leaves of Phaseolus vulgaris, the extractable activity of adenosine 5-phosphosulfate sulfotransferase increased ten-fold. When chloroplast development took place in air enriched with 3.5 l H₂S·l^-1 there was a decrease in adenosine 5-phosphosulfate sulfotransferase activity. Cyst(e)ine in concentrations up to 1 mM (in the external medium) did not affect the increase in adenosine 5-phosphosulfate sulfotransferase activity in intact plants. In plants with excised roots, 0.75 mM cyst(e)ine inhibited this increase. In green primary leaves, H₂S or cyst(e)ine treatment resulted in a decrease of extractable adenosine 5-phosphosulfate sulfotransferase activity. In intact plants, this effect of cyst(e)ine was observed at a concentration of 1 mM, and in plants with excised roots, 0.25 mM had a comparable effect.In developing plants, the extractable activities of O-acetyl-L-serine sulfhydrylase (EC 4.2.99.9) and ribulosebisphosphate carboxylase (EC 4.1.1.39.) were not affected by H₂S or cyst(e)ine.Abbreviations APS adenosine 5-phosphosulfate - APSSTase adenosine 5phosphosulfate sulfotransferase - BSA bovine serum albumin - DTE dithioerythritol - EDTA ethylenediaminetetra-acetic acid - OASSase O-acetyl-L-serine sulfhydrylase - PAPS adenosine 3-phosphate 5-phosphosulfate - POPOP 1,4 Di 2-(5-phenyloxazolyl)-benzene - PPO 2,5-diphenyloxazol - RubP ribulose-bisphosphate - RubPCase ribulosebiphosphate carboxylase This is no. 8 in the series Regulation of Sulfate Assimilation in Plants. The term cysteine is used when it is clear that cystine is not involved; cyst(e)ine is used for an undefined mixture of cysteine and cystine. The concentrations are expressed in all cases relative to cysteine     

New applications of ruthenium solar cell sensitizers N3 and N719 as luminescence turn-on anion sensors

Optical sensing of F⁻, Cl⁻, Br⁻, I⁻, OAc⁻, , , and by cis-dithiocyanatobis(2,2′-bipyridyl-4,4′-dicarboxylic acid)ruthenium(II) (N3) and bis(tetrabutylammonium) cis-dithiocyanatobis(2,2′-bipyridine-4-COOH,4′-COO⁻)ruthenium(II) (N719) have been investigated in dimethyl sulfoxide (DMSO), by means of UV-Vis absorption and emission spectrophotometric titrations. Additions of F⁻, OAc⁻, and in DMSO solution caused obvious UV-Vis spectral changes with appearance of several isosbestic points, and remarkable emission enhancements along with large blue shifts in emission bands. The values of F⁻-induced emission intensity enhancement factor (emission quantum yield enhancement factor), I/I₀ (φ/φ₀), were found to be 40 (86) and 38 (58) for N3 and N719, respectively. No obvious spectral changes were observed upon addition of Cl⁻, Br⁻, I⁻, and in DMSO solutions. Luminescent F⁻ sensing in DMSO/H₂O (4:1, v/v) has also been demonstrated to be operative with a luminescence enhancement factor of 12, indicating that N3 is very potential for practical application as fluorescent anion sensor in aqueous solution. An interaction mechanism of anion-induced deprotonation of N3 and N719 was confirmed, and the deprotonation reaction equilibrium constants of N3 and N719 were derived as well.     

Structure and differentiation of the cell wall of Phytophthora palmivora: Cysts,hyphae and sporangia

Summary Walls from cysts, hyphae and sporangia of Phytophthora palmivora consist chiefly (ca. 90% dry wt) of -glucans with 1,3-, 1,4- and 1,6-links. The glucans are predominatly -1,3-linked but there are significant differences in the relative proportion of 1,3-, 1,6- and 1,4-linked glucosyl residues among the three wall types. There are also differences in protein content, susceptibility to degradation by various -glucanases, and surface texture. The isolated cyst wall consists solely of a thin fabric of long, tightly interwoven, randomly oriented microfibrils. Both inner and outer surfaces of the cyst wall are distinctly microfibrillar. The hyphal wall has two different textures; the internal surface is distinctly microfibrillar while the external surface is non-fibrillar. In a germinated cyst, there is a zone of demarcation where the microfibrils of the cyst wall disappear into the smooth outer texture of the germ tube wall. An exo--1,3-glucanase preferentially removed the amorphous material of the outer surface of the germ tube leaving exposed a continuous microfibrillar fabric from cyst to hyphal tube. Conceivably, the textural and structural differentiation of the cell wall may play a decisive role in cellular morphogenesis.     

Synthesis, photophysical and electrochemical properties, and biological labelling studies of luminescent cyclometallated iridium(III) bipyridine-aldehyde complexes

We report the synthesis, characterisation, and photophysical and electrochemical properties of a series of luminescent cyclometallated iridium(III) bipyridine-aldehyde complexes [Ir(N-C)₂(bpy-CHO)](PF₆) (HN-C=2-phenylpyridine, Hppy (1); 2-(4-methylphenyl)pyridine, Hmppy (2); 1-phenylpyrazole, Hppz (3); 3-methyl-1-phenylpyrazole, Hmppz (4); 7,8-benzoquinoline, Hbzq (5); 2-phenylquinoline, Hpq (6); bpy-CHO=4-formyl-4^′-methyl-2,2^′-bipyridine). The X-ray crystal structures of complexes 1 and 4 have been determined. On the basis of the photophysical data, the emission of these complexes is assigned to an excited state of predominantly triplet metal-to-ligand charge-transfer (³MLCT) (dπ(Ir) → (bpy-CHO)) character. For complex 6, the excited state is also mixed with substantial (³IL) () (pq⁻) character. The protein bovine serum albumin has been labelled with these complexes to produce luminescent bioconjugates. The photophysical properties of the luminescent conjugates have also been investigated.     

Synthesis and single crystal structural investigations on quaternary salts of tellurated alkylamine derivatives

Tellurated alkylamine derivatives , , and have been synthesized by reacting appropriate organic halides with the nucleophile 4-CH₃OC₆H₄Te⁻ or Te²⁻ generated in situ by borohydride reduction of (4-CH₃OC₆H₄Te)₂ or Te powder followed by reaction with HCl of appropriate concentration. The zwitterionic species was generated when single crystals of 2 were grown in methanol at 0 °C. Complexes 1-4 exhibit characteristic ¹H NMR spectra. The single crystal structures of 1-4 and 2a have been determined. In the crystals of 1, C-H?π distances have been found to be 3.31(7)-3.59(5) Å. In both 2 and 2a, weak Te?Cl interactions (3.54(2) -3.62(2) Å) are observed. The C-H?π distance in the crystal of 2 is 3.19(0) Å. In 2a and 3, water hydrogen bonds connect the water molecules with the end groups from different molecules. In the case of 3, Te?Cl weak interactions involving the Cl⁻ ions connect together two such chains. The geometry of Te in 1 is V shaped. In 2 and 3 it is pseudo trigonal bipyramidal, and in 2a, it is square pyramidal. However, in the latter case it becomes distorted octahedral due to weak Te?Cl secondary interactions. The geometry about Te in 4 is distorted octahedral due to weak Te?Cl interactions involving Cl⁻ ions. However, there are no intermolecular Te?Cl interactions.     

Catalytic hydroxylation of 1-propanol by platinum NCN and PCP pincer complexes using CuCl2 as oxidant

The novel PCP-pincer Pt(II) complex, has been prepared and characterized by ¹H, ³¹P, and ¹³C NMR spectroscopy. The molecular structure of 1 has been determined through a single-crystal X-ray diffraction study. The pincer-ligated platinum complexes 1 and PtCl{C₆H₃-2,6-(CH₂NEt₂)₂} (2) have been explored as catalysts for the hydroxylation of 1-propanol to 1,3-propanediol under mild conditions. Product ratios and turnover numbers achieved with both complexes compare favorably to those obtained with [PtCl₄]²⁻. Moreover, the pincer complexes catalyze this transformation even upon replacement of PtCl₄ by the more economical CuCl₂ as the requisite stoichiometric oxidant. Analysis of the reaction mixture by ³¹P NMR spectroscopy following the hydroxylation of 1-propanol by 1 in the presence of CuCl₂ revealed that 1 is partially converted to the ring substituted complex, . The molecular structure of 3 has been determined through a single-crystal X-ray diffraction study.     

Sequence divergences between cyst nematode effector protein orthologs may contribute to host specificity

The soybean cyst nematode (Heterodera glycines) and the closely related sugar beet cyst nematode (Heterodera schachtii) are devastating pathogens of plant roots that use secreted effector proteins to engage in sophisticated host-parasite interactions. While H. schachtii infects and reproduces readily on the roots of Arabidopsis thaliana, H. glycines rarely is able to infect this model plant. The molecular basis for differing host ranges remains obscure but likely involves differences between nematode effector proteins and the recognition of host factors. Recently we reported that constitutive expression of the H. schachtii 10A06 effector protein gene (Hs-10A06) in Arabidopsis affected plant morphology and increased susceptibility to H. schachtii and that the 10A06 protein functions through its interaction with Arabidopsis spermidine synthase 2 (SPDS2). Therefore, we investigated whether differences between cyst nematode effector protein orthologs in two nematode species have a role in mediating host specificity. Here, we show that, similar to Hs-10A06, ectopic expression of H. glycines 10A06 (Hg-10A06) in Arabidopsis affected leaf number and root length, however, to a much lesser extent. More importantly, no effect of Hg-10A06 overexpression on Arabidopsis susceptibility to H. schachtii was observed. While we found that Hg-10A06 can weakly interact with Arabidopsis SPDS2 in yeast-two hybrid assays, this ability to interact with SPDS2 was decreased approximately five-fold compared with Hs-10A06. Collectively, these data suggest that sequence divergence between cyst nematode effector protein orthologs could contribute in determining cyst nematode host range.Key words: Heterodera schachtii, arabidopsis, 10A06 effector protein, spermidine synthase 2Cyst nematodes are sedentary pathogens of roots of many economically important crop plants and induce the formation of specialized feeding cells, so-called syncytia, that provide the nematodes with nourishment. The infection process is mediated through secretion of an array of nematode effector proteins inside plant tissues and cells. One of these effector proteins is 10A06, which was initially identified from a gland cell cDNA library from H. glycines, the soybean cyst nematode.¹ The 927 bp full-length H. glycines Hg-10A06 cDNA (GenBank Accession {"type":"entrez-nucleotide","attrs":{"text":"AF502391","term_id":"30315231","term_text":"AF502391"}}AF502391) encoded a predicted protein of 308 amino acids with an N-terminal signal peptide of 17 amino acids for secretion. Recently, we identified the orthologous 10A06 sequence from the sugar beet cyst nematode H. schachtii (Hs-10A06), which is able to infect the model plant Arabidopsis thaliana. The Hs-10A06 cDNA (GenBank Accession {"type":"entrez-nucleotide","attrs":{"text":"GQ373256","term_id":"255528984","term_text":"GQ373256"}}GQ373256) contained an open reading frame of 858 bp encoding a 285-amino acid protein with an N-terminal signal peptide for secretion.² Sequence alignment of H. glycines and H. schachtii 10A06 proteins revealed a strong homology between both orthologues with 86% identity and 87% similarity. The largest difference between the two proteins is the lack of a stretch of 23 amino acids in Hs-10A06. Additionally, a region of 15 amino acid residues located between amino acid 167 and 181 exhibited a high degree of divergence between both proteins. Constitutive expression of Hs-10A06 in Arabidopsis affected plant morphology and increased susceptibility to H. schachtii.² We uncovered in yeast two-hybrid assays that the Hs-10A06 protein interacts with Arabidopsis SPDS2, a key enzyme involved in polyamine biosynthesis, to mediate susceptibility. Here, we assessed the effects of ectopic Hg-10A06 expression in the non-host Arabidopsis on plant morphology and nematode susceptibility. Moreover, we assayed whether Hg-10A06 also is able to interact with SPDS2 from Arabidopsis.     

Substitution, abstraction and addition chemistry of low-coordinate gallium and indium ligand systems

Substitution, abstraction and addition processes have been shown to be viable chemistries for the modification of ligand systems featuring heavier group 13 element donor atoms. Thus substitution of the bromide in Cp^∗Fe(CO)₂In(Br)Mes^∗ (1) can be carried out with retention of the Fe-In bond, using 1 equiv. of the aryloxide nucleophile [OC₆H₄^tBu-4]⁻ to give Cp^∗Fe(CO)₂In(OC₆H₄^tBu-4)Mes^∗ (2). Structural and spectroscopic comparisons of 1 and 2 reveal that variation in the steric and/or π donor properties of the indyl ligand substituents have little effect on the nature of the Fe-In bond. Sequential reaction of [Cp^∗Fe(CO)₂]₂GaCl (3) with the halide abstraction agent and 4-picoline in dichloromethane proceeds via the known two-coordinate gallium cation (4). The net result is replacement of the gallium bound chloride substituent with a 4-picoline moiety, yielding (5) via a two-step abstraction/addition process. 5 represents only the second structurally characterized complex containing a cationic three-coordinate gallium centre, and the first displaying bonds to a transition metal.     

Protein phosphorylation in the facultative chemolithotrophThiobacillus novellus

The phosphorylation of at least five proteins with M_r of about 160,000; 93,000; 85,000; 45,000; and 29,000 respectively was demonstrated in crude extracts from the facultative chemolithotrophThiobacillus novellus. The incorporation of [-³²P]phosphate from ATP into these proteins was dependent on the presence of magnesium ion. The phosphorylation reactions were found to be reversible and required 12.5 mM NaF for maximal activity, indicating the action of phosphatases. In addition, 3,5-cAMP had little effect on protein kinase activity, whereas Ca²⁺ alone was weakly stimulatory. This activation was enhanced by the addition of 3,5-cAMP. Ca²⁺ with calmodulin had a strong stimulatory effect on phosphate incorporation into the proteins. A highly purified preparation containing only the 160, 93, and 85 kDa proteins phosphorylated histone, whereas the uptake of³²P by the three proteins was inhibited. Rabbit muscle phosphorylase b prevented incorporation of radiolabel only into the 160 and 93 kDa proteins.     

Oxomolybdenum(III, IV and V) complexes of thiofunctional ligands

Reaction of [MoO₂(acac)₂] with (S is a thioether, S′ a thiophenolate function) yielded the compound Li₇(thf)₁₇{MoO}₈ · 10thf · hexane, where {MoO}₈ represents one 1, three (2, linked, via the oxo group, to [Li(thf)₃]⁺) and two (3a, linked by two [Li(thf)₂]⁺).A mixed-valent variant of 3, (3b, with an additional[Li(thf)₃]⁺ attached to S′), was also identified. The compounds model features pertinent to oxo-transferases containing the molybdopterin cofactor.     

The use of heteronuclear cross-polarization for backbone assignment of 2H-, 15N- and 13C-labeled proteins: A pulse scheme for triple-resonance 4D correlation of sequential amide protons and 15N

Summary A new four-dimensional pulse scheme is described for the main-chain assignment of proteins by means of the J connectivity of the amide proton and nitrogen resonances of adjacent residues. Since the new experiment, 4D CP-HN(COCA)NH, involves heteronuclear cross-polarization for magnetization transfer from ¹³C=O to ¹⁵N via ¹³C, a relatively strong WALTZ-16 decoupling rf field is applied to ¹³C during magnetization transfer. Consequently, ¹³C is effectively decoupled from its attached ²H in the case of deuterated proteins, in the absence of a decoupling rf field for ²H. This efficiently improves the sensitivity of the experiment through ¹³C line narrowing. The experiment was performed on a randomly 60% deuterated protein, and the sensitivity of the final 4D spectrum was found to be excellent.