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PubMed2006

Sildenafil citrate (Viagra) complexes with bivalent ions.

The interaction of Ca(2+)-ions with sildenafil citrate (Viagra) leads to the precipitation of a new polymorph variety of sildenafil base. Under the same conditions, Mg(2+), Zn(2+), and Cd(2+) ions form structurally related crystalline complexes of the composition Me(2+)C(28)H(34)N(6)O(11)S. Lattice parameters have been determined showing that magnesium compound belongs to an orthorhombic system, while the zinc and cadmium compounds are its monoclinic distortions. All three compounds are thermally stable, undergoing decomposition above 175 degrees C with the consequent formation of carbonates Me(2+)CO(3) and oxides.

Cations, Divalent↗
PubMed1999

Tandem affinity tags for the purification of bivalent anti-DNA single-chain Fv expressed in Escherichia coli.

Antibodies to DNA define an important autospecificity that arises in systemic lupus erythematosus (SLE). To elucidate the molecular features that may explain the pathogenesis of SLE, a heterologous system for expression of cloned V genes is often desirable. Here, a single-chain Fv coding domain was constructed by using the heavy- and light-chain V genes of a high-affinity site-directed mutant of the murine anti-dsDNA autoantibody, 3H9. This scFv was joined in frame to the c-jun leucine zipper for dimerization, and to two affinity tags, domain B of the staphylococcal protein A and a pentahistidine peptide, for purification. Dimerization of the scFv was determined by size-exclusion chromatography. The yields of the scFv following affinity purification on IgG agarose or Ni-NTA agarose were compared, and the activities of the resulting protein fractions were determined. A two-step purification of periplasmic extracts on Ni-NTA agarose and IgG agarose, followed by elution with 3.5 M MgCl(2), yielded scFv with the highest specific activity. The final purified material bound DNA by ELISA, electrophoretic mobility shift assay, and immunofluorescence of fixed Hep-2 cells. Antibodies purified in this fashion should have applications in structure/function studies in which it is essential to generate highly purified antigen-combining sites.

Affinity Labels↗
PubMed1987

Binding sites for horseradish peroxidase on the cell surface. Suppression of binding by gangliosides and effects of some bivalent cations.

The cytochemical reaction for surface-bound horseradish peroxidase (HRP) on cultured HeLa cells, GH3 cells, and isolated rat liver cells was suppressed by 30 microM monosialoganglioside, by 30 microM trisialoganglioside, or by 5 mM CMP-neuraminic acid. The reaction was also suppressed by 10 mM chitotriose or by 10 mM UDP-galactose, a galactose acceptor and donor, respectively, for galactosyl-transferase. The addition of 2 mM Mn2+ to the incubation medium with HRP suppressed the reaction for surface-bound HRP, and the addition of 10-20 mM Ca2+ intensified the reaction. The addition of 2 mM Zn2+ caused less inhibition than that of 2 mM Mn2+, and the addition of 2 mM Co2+ caused either a slight inhibition, or no inhibition. These observations support the hypothesis that HRP may be bound to a glycosyltransferase at the cell surface.

Animals↗
PubMed1975

[The mechanism of antigen-antibody reaction at the basement membrane zone in bullous pemphigoid. The question of interaction of complement factors and bivalent cations (author's transl)].

1. In a medium poorly cationic indirect immunfluorescence of the basement membrane zone normally observed with preparations of sera from patients with pemphigoid has not be seen. 2. The fluorescence of basement membrane zone takes place if Fe2+ ions are added together with the antibasement zone antibodies. Evidently Fe2+ ions are essential for fixing antibodies. 3. With the method of indirect fluorescence the C3 component of complement could be observed after fixation of pemphigoid antibodies to the basement membrane zone. 4. EDTA prevents binding C3 too. Only after adding Fe2+ (not with Ca2+ or Mg2+) complement IF staining is demonstrable. Apparently EDTA causes the loss of fixing antibasement zone antibodies at first. Thereafter complement staining reaction is negative. 5. Participation of C1 could not be demonstrated using indirect IF staining. The alternate pathway to activation of C3 is discussed.

Antigen-Antibody Reactions↗
PubMed1995

High-resolution cytological localization of the XhoI and EcoRI repeat sequences in the pachytene ZW bivalent of the chicken.

The pachytene ZW pair of the chicken has been studied with a novel combination of fluorescent in situ hybridization and silver staining of the synaptonemal complexes, and with electron microscopic in situ hybridization. Probes for the EcoRI and the XhoI repeat sequences were used for light microscopy and probes for XhoI for electron microscopy. XhoI repeats are pericentromeric and correspond to 27.3% of the W axis length. EcoRI repeats form three distinct domains: domain I covers the distal 19.4% of the late-synapsing arm, and domains II and III cover respectively 21.7% and 8.5% of the W axis. The early-synapsing end containing the recombination nodule is free from any of both signals. This non-hybridizing region accounts for 25.9% of the W axis. It is suggested that this region is composed of a proximal region containing other repeat types and a terminal region which is the recombining or pseudoautosomal region. The successful combination of silver staining and fluorescent in situ hybridization will be generally useful for high-resolution localization of DNA sequences in meiotic chromosomes.

Animals↗
PubMed1983

A central mechanism of action for taurine: osmoregulation, bivalent cations, and excitation threshold.

A postulated zinc-taurine complex, with a zinc affinity intermediate between that for glutamic acid dehydrogenase and the calcium binding protein(s), provides an explanation for a series of seemingly unrelated biochemical and physiological effects of taurine. The proposed complex suggests a central mechanism for the action of taurine, such as a bicarbonate and pH dependent influence on calcium and zinc movements (and vice versa), the osmoregulatory role of taurine, and its effect on the excitation threshold.

Animals↗