In situ preparation of protein-"smart" polymer conjugates with retention of bioactivity

Protein-polymer conjugates are widely used in biotechnology and medicine, and new methods to prepare the bioconjugates would be advantageous for these applications. In this report, we demonstrate that bioactive "smart" polymer conjugates can be synthesized by polymerizing from defined initiation sites on proteins, thus preparing the polymer conjugates in situ. In particular, free cysteines, Cys-34 of bovine serum albumin (BSA) and Cys-131 of T4 lysozyme V131C, were modified with initiators for atom transfer radical polymerization (ATRP) either through a reversible disulfide linkage or irreversible bond by reaction with pyridyl disulfide- and maleimide-functionalized initiators, respectively. Initiator conjugation was verified by electrospray-ionization mass spectroscopy (ESI-MS), and the location of the modification was confirmed by muLC-MSMS (tandem mass spectrometry) analysis of the trypsin-digested protein macroinitiators. Polymerization of N-isopropylacrylamide (NIPAAm) from the protein macroinitiators resulted in thermosensitive BSA-polyNIPAAm and lysozyme-polyNIPAAm in greater than 65% yield. The resultant conjugates were characterized by gel electrophoresis and size exclusion chromatography (SEC) and easily purified by preparative SEC. The identity of polymer isolated from the BSA conjugate was confirmed by (1)H NMR, and the polydispersity index was determined by gel permeation chromatography (GPC) to be as low as 1.34. Lytic activities of the lysozyme conjugates were determined by two standard assays and compared to that of the unmodified enzyme prior to polymerization; no statistical differences in bioactivity were observed.     

Photokatalytische Systeme. XLIV. Über das Intervalence-Charge-Transfer-Verhalten von Ionenpaarassoziaten des Octacyanomolybdats

Photocatalytic Systems. XLIV. On the Intervalence Charge Transfer Behaviour of Ion Pairs of Octacyanomolybdate [Mo(CN)₈]^4? ions are forming coloured ion pairs with Fe^III, Cu^II, U^VIO₂, and V^IVO, the long-wavelength absorption band of which could be assigned to an intervalence charge-transfer transition. From the solvent dependence of the IT band, interionic distances could be estimated.     

GnRH binding RNA and DNA Spiegelmers: a novel approach toward GnRH antagonism

Mirror-image oligonucleotide ligands (Spiegelmers) that bind to the pharmacologically relevant target gonadotropin-releasing hormone I (GnRH) with high affinity and high specificity have been identified using the Spiegelmer technology. GnRH is a decapeptide that plays an important role in mammalian reproduction and sexual maturation and is associated with several benign and malignant diseases. First, aptamers that bind to D-GnRH with dissociation constants of 50-100 nM were isolated out of RNA and DNA libraries. The respective enantiomers of the DNA and RNA aptamers were synthesized, and their binding to L-GnRH was shown. These Spiegelmers bind to L-GnRH with similar affinity to that of the corresponding aptamers that bind to D-GnRH. We further demonstrated dose-dependent inhibition of GnRH-induced Ca(2+) release in Chinese hamster ovary cells that were stably transfected with the human GnRH receptor.     

The number of kekulé structures for rectangle-shaped benzenoids,part VIII: Some perforated rectangles

Summary Classes of coronoids (or degenerate coronoids), r and q are obtained by providing the rectangle-shaped benzenoids R^j (m, 3) with a naphthalenic or pyrenic hole, respectively. The numbers of Kekulé structures (K) are studied. It was found for theK numbers of the classes in question:r=4/5R andq=1/5R. The classes r and q are similar to r and q, respectively, but the naphthalenic or pyrenic hole is oriented in a different way. For these classes it was found:r=3/5R,q=2/5R.

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Die Anzahl von Kekulé-Strukturen für hochkondensierte Benzenoide mit rechteckigem Umriß, 8. Mitt.: Einige perforierte benzenoide Rechteck-Strukturen

Zusammenfassung Es werden Klassen r und q von Coronoiden (oder degenerierten Coronoiden) erhalten, wobei Benzenoide mit rechteckigem Umriß, R^j(m, 3), mit einem Naphthalin- oder Pyren-Loch versehen werden. Die Anzahl von Kekulé-Strukturen (K) wird untersucht; dabei wurde für die untersuchten Klassenr=4/5R undq=1/5R gefunden. Die Klassen r und q verhalten sich ähnlich, das Naphthalin- oder Pyren-Loch ist jedoch anders orientiert; dafür wurder=3/5R undq=2/5R gefunden.

     

Bi- and trinuclear oxalamidinate complexes of palladium as catalysts in the copper-free Sonogashira reaction and in the Negishi reaction

The binuclear complex [(acac)Pd(oxam)Pd(acac)] 1 (oxam: tetraphenyl oxalic amidinate) has been prepared from H₂oxam and Pd(acac)₂ in excellent yield. The complex was characterized by elemental analyses, mass spectroscopy, ¹H NMR, ¹³C NMR spectroscopy and in the solid state by X-ray single crystal diffraction analyses. 1 consists of a bimetallic centrosymmetric unit in which the planar oxam ligand acts in a bis-chelating fashion. Each palladium center is in a planar environment.The complex 1 acts as highly selective pre-catalyst in the copper-free Sonogashira reaction between 4-bromoacetophenone and phenylacetylene. Its long-time catalytic activity is higher than that of the related binuclear complex 2 (oxam: tetra-p-tolyl oxalic amidinate) or that of the trinuclear compound [(acac)Pd(oxam)Zn(oxam)Pd(acac)] (3), the solid-state structure of which was also determined by an X-ray structural analysis of single crystals. In addition, 2 is an active and extremely selective pre-catalyst for the Negishi reaction between 3,5,6,8-tetrabromophenanthroline and R-CC-ZnCl (R: Ph, (ⁱprop)₃Si) to form tetra-alkyne-substituted derivatives.     

Nature of mechanically induced defects in polycrystalline hydrargillite and Al2O3-A1(OH)3 studied by radiation methods

Mechanical treatment of hydrargillite Al(OH)₃ in a vibrational grinder and in a ball-mill grinder increases the dispersion, the specific surface area, and the unit-cell constants. An x-ray amorphous phase forms. Differences in the nature and in the concentration of paramagnetic centers arise on irradiation (77 K) of the mechanically treated samples. Anion vacancies and oxygen ions with disrupted short-range order appear in the mechanically treated samples. Their concentration increases with increased time of mechanical treatment. Pulsed thermal treatment of hydrargillite forms the Al₂O₃-Al(OH)₃ system and anneals the mechanically induced defects.Translated from Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, No. 7, pp. 1463–1467, July, 1990.     

Phase stability in the systems AeAl(2-x)Mgx (Ae = Ca, Sr, Ba): electron concentration and size controlled variations on the laves phase structural theme

The systems AeAl(2-x)Mgx (Ae = Ca, Sr, Ba) display electron concentration induced Laves phase structural changes. However, the complete sequence MgCu2 --> MgNi2 --> MgZn2 with increasing x (decreasing electron count) is only observed for Ae = Ca. Compounds SrAl(2-x)Mgx (0 < x < or = 2) and BaAl(2-x)Mgx (x = 0.85 and 2.0) were synthesized and structurally characterized by X-ray diffraction experiments. For the Sr system the structural sequence CeCu2 --> MgNi2 --> MgZn2 occurs with increasing Mg content x. Thus, larger Sr does not allow the realization of the MgCu2 structure at low x. For Ae = Ba a binary compound BaAl2 does not exist, but more Ba-rich Ba7Al13 forms. The reinvestigation of the crystal structure of Ba7Al13 by selected area and convergent beam electron diffraction in a transmission electron microscope revealed a superstructure, which subsequently could be refined from single X-ray diffraction data. The formula unit of the superstructure is Ba21Al40 (space group P31m, Z = 1, a = 10.568(1) angstroms, c = 17.205(6) angstroms). In Ba21Al40 a size match problem between Ba and Al present in Ba7Al13 is resolved. The structure of Ba7Al13 (Ba21Al40) can be considered as a Ba excess variant of the hexagonal MgNi2 Laves phase type structure. An incommensurately modulated variant of the MgNi2 structure is obtained for phases BaAl(2-x)Mgx with x = 0.8-1. At even higher Mg concentrations a structural change to the proper MgZn2 type structure takes place.     

Dibenzotetraaza crown ethers: a new family of crown ethers based on o-phenylenediamine

Dibenzotetraaza (DBTA) crown ethers possess two o-phenylenediamine moieties. They are homologues of dibenzo crown ether phase-transfer catalysts and were prepared from the condensation of benzimidazoles with oligo(ethyleneglycol) dichlorides and oligo(ethyleneglycol) ditosylates. Compounds with ring sizes ranging from 18-crown-6 to 42-crown-14 were prepared. In addition, various altered benzimidizoles were used to produce DBTA crown ethers with modified substituents and ether bridges, as well as benzimidazolidine crown ethers. The synthetic approach presented here proved to be a convenient route to a new family of crown ethers with overall yields of up to 48% based on the benzimidazole. Yields for the ring-closing step were generally high, ranging from 51% to 94%, without the need for high-dilution conditions. Reaction of the DBTA crown ethers with alkyl and benzyl halides was found to be a facile way to obtain the corresponding tetra(N-organyl) compounds. Picrate extraction studies were carried out to determine phase-transfer catalytic capabilities. Extraction efficiencies for alkali-metal ions were lower than those for dibenzo-18-crown-6. Efficiencies were higher for other metal ions, with some selectivity for Pb(2+). Tetra(N-methyl) DBTA-18-crown-6 generally exhibited higher extraction efficiencies than its N-H analogue, but the selectivity was lower.