The identification of 14 alpha,17 beta-dihydroxyandrost-4-en-3-one monohydrate and 14 alpha,17 beta-dihydroxyandrosta-1,4-dien-3-one monohydrate, metabolites of androstenedione in Mucor piriformis

The microorganism Mucor piriformis transforms androst-4-ene-3,17-dione into a major and several minor metabolites. X-ray crystallographic analysis of two of these metabolites was undertaken to determine unambiguously their composition and chirality. Crystals belong to the orthorhombic space-group P2(1)2(1)2(1), with a = 7.199(4) angstrom and a = 6.023(3) angstrom, b = 11.719(3) angstrom and b = 13.455(4) angstrom, c = 20.409(3) angstrom and c = 20.702(4) angstrom for the two title compounds, respectively. The structures have been refined to final R values of 0.060 and 0.040, respectively.

Conformational studies on beta-bend containing a cis peptide unit

Conformational studies have been carried out on the X-cis-Pro tripeptide system (a system of three linked peptide units, in the trans-cis-trans configuration) using energy minimization techniques. For X, residues Gly, L-Ala, D-Ala and L-Pro have been used. The energy minima have been classified into different groups based upon the conformational similarity. There are 15, 20, 18 and 6 minima that are possible for the four cases respectively and these fall into 11 different groups. A study of these minima shows that, (i) some minima contain hydrogen bonds - either 4-->1 or 1-->2 type, (ii) the low energy minima qualify themselves as bend conformations, (iii) cis' and trans' conformations are possible for the prolyl residue as also the C(gamma)-endo and C(gamma)-exo puckerings, and (iv) for Pro-cis-Pro, cis' at the first prolyl residue is ruled out, due to the high energy. The available crystal structure data on proteins and peptides, containing cis-Pro segment have been examined with a view to find the minima that occur in solid state. The data from protein show that they fall under two groups. The conformation at X in X-cis-Pro is near extended when it is a non-glycyl residue. In both peptides and proteins there exists a preference for trans' conformation at prolyl residue over cis' when X is a non-glycyl residue. The minima obtained can be useful in modelling studies.

Differential stability of beta-sheets and alpha-helices in beta-lactamase: a high temperature molecular dynamics study of unfolding intermediates

Beta-Lactamase, which catalyzes beta-lactam antibiotics, is prototypical of large alpha/beta proteins with a scaffolding formed by strong noncovalent interactions. Experimentally, the enzyme is well characterized, and intermediates that are slightly less compact and having nearly the same content of secondary structure have been identified in the folding pathway. In the present study, high temperature molecular dynamics simulations have been carried out on the native enzyme in solution. Analysis of these results in terms of root mean square fluctuations in cartesian and [phi, psi] space, backbone dihedral angles and secondary structural hydrogen bonds forms the basis for an investigation of the topology of partially unfolded states of beta-lactamase. A differential stability has been observed for alpha-helices and beta-sheets upon thermal denaturation to putative unfolding intermediates. These observations contribute to an understanding of the folding/unfolding processes of beta-lactamases in particular, and other alpha/beta proteins in general.

Conformational analysis and design of cross-strand disulfides in antiparallel beta-sheets

Cross-strand disulfides bridge two cysteines in a registered pair of antiparallel beta-strands. A nonredundant data set comprising 5025 polypeptides containing 2311 disulfides was used to study cross-strand disulfides. Seventy-six cross-strand disulfides were found of which 75 and 1 occurred at non-hydrogen-bonded (NHB) and hydrogen-bonded (HB) registered pairs, respectively. Conformational analysis and modeling studies demonstrated that disulfide formation at HB pairs necessarily requires an extremely rare and positive chi(1) value for at least one of the cysteine residues. Disulfides at HB positions also have more unfavorable steric repulsion with the main chain. Thirteen pairs of disulfides were introduced in NHB and HB pairs in four model proteins: leucine binding protein (LBP), leucine, isoleucine, valine binding protein (LIVBP), maltose binding protein (MBP), and Top7. All mutants LIVBP T247C V331C showed disulfide formation either on purification, or on treatment with oxidants. Protein stability in both oxidized and reduced states of all mutants was measured. Relative to wild type, LBP and MBP mutants were destabilized with respect to chemical denaturation, although the sole exposed NHB LBP mutant showed an increase of 3.1 degrees C in T-m. All Top7 mutants were characterized for stability through guanidinium thiocyanate chemical denaturation. Both exposed and two of the three buried NHB mutants were appreciably stabilized. All four HB Top7 mutants were destabilized (Delta Delta G(0) = -3.3 to -6.7 kcal/mol). The data demonstrate that introduction of cross-strand disulfides at exposed NHB pairs is a robust method of improving protein stability. All four exposed Top7 disulfide mutants showed mild redox activity. Proteins 2011; 79: 244-260. (C) 2010 Wiley-Liss, Inc.

Formation of a single, rotated-Brass (1 1 0)< 5 5 6 > texture by hot cross-rolling of an Al-Zn-Mg-Cu-Zr alloy

The present work describes the evolution of a strong, single-component rotated-Brass ((1 1 0) < 5 5 6 >) texture in an Al-Zn-Mg-Cu-Zr alloy by an uneven hot cross-rolling with frequent interpass annealing. This texture development is unique because hot rolling of aluminum alloys results in orientation distribution along the ``beta-fibre''. It has been demonstrated that the deformation by cross-rolling of a partially recrystallized grain structure having rotated-Cube and Goss orientations, and the recrystallization resistance of near-Brass-oriented elongated grains play a critical role in development of this texture. (C) 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

AB-initio study of alpha-silyl and beta-silyl substituent effects on vinyl radicals

The geometries of alpha- and beta-silyl substituted vinyl radicals and of alpha,beta-disilylvinyl radical have been optimised with the STO-3G and the STO-3G* basis sets. The relative stabilities of various conformers have been determined at the UMP2/6-31G* level. The stabilisation of vinyl radicals through alpha-silyl substitution is larger than that due to corresponding alkyl groups. The presence of an alpha-silyl group also leads to a tendency towards linearisation of the vinyl radical centre and a corresponding reduction in the inversion barrier. In marked contrast, the beta-silyl effect is negligible. The geometric, conformational and energetic consequences are insignificant. Overall, the silyl substituent effect at vinyl radicals is very different from that computed earlier for the vinyl cations, but qualitatively similar to that found in carbanions.

Additive Spanners and (alpha, beta)-Spanners

An (alpha, beta)-spanner of an unweighted graph G is a subgraph H that distorts distances in G up to a multiplicative factor of a and an additive term beta. It is well known that any graph contains a (multiplicative) (2k - 1, 0)-spanner of size O(n(1+1/k)) and an (additive) (1, 2)-spanner of size O(n(3/2)). However no other additive spanners are known to exist. In this article we develop a couple of new techniques for constructing (alpha, beta)-spanners. Our first result is an additive (1, 6)-spanner of size O(n(4/3)). The construction algorithm can be understood as an economical agent that assigns costs and values to paths in the graph, purchasing affordable paths and ignoring expensive ones, which are intuitively well approximated by paths already purchased. We show that this path buying algorithm can be parameterized in different ways to yield other sparseness-distortion tradeoffs. Our second result addresses the problem of which (alpha, beta)-spanners can be computed efficiently, ideally in linear time. We show that, for any k, a (k, k - 1)-spanner with size O(kn(1+1/k)) can be found in linear time, and, further, that in a distributed network the algorithm terminates in a constant number of rounds. Previous spanner constructions with similar performance had roughly twice the multiplicative distortion.

Characterization of the DNA-binding domain of beta protein, a component of phage lambda Red-pathway by UV catalyzed cross-linking

beta protein, a key component of Red-pathway of phage lambda is necessary for its growth and general genetic recombination in recombination-deficient mutants of Escherichia coli. To facilitate studies on structure-function relationships, we overexpressed beta protein and purified it to homogeneity. A chemical cross-linking reagent, glutaraldehyde, was used to stabilize the physical association of beta protein in solution. A 67-kDa band, corresponding to homodimer, was identified after separation by SDS-polyacrylamide gel electrophoresis. Stoichiometric measurements indicated a site-size of 1 monomer of beta protein/5 nucleotide residues. Electrophoretic gel mobility shift assays suggested that beta protein formed stable nucleoprotein complexes with 36-mer, but not with 27- or 17-mer DNA. Interestingly, the interaction of beta protein with DNA and the stability of nucleoprotein complexes was dependent on the presence of MgCl2, and the binding was abolished by 250 mM NaCl. The K-d of beta protein binding to 36-mer DNA was on the order of 1.8 x 10(-6) M. Photochemical cross-linking of native beta protein or its fragments, generated by chymotrypsin, to 36-mer DNA was performed to identify its DNA-binding domain. Characterization of the cross-linked peptide disclosed that amino acids required for DNA-binding specificity resided within a 20-kDa peptide at the N-terminal end. These findings provide a basis for further understanding oi the structure and function of beta protein.

Chemo- and stereoselective reduction of alpha,beta-epoxyketones with diisopropoxytitanium(III) tetrahydroborate

Reduction of alpha,beta-epoxyketones with diisopropoxytitanium(III) tetrahydroborate in dichloromethane under mild conditions (-78 degrees--> -20 degrees C) provides anti- (or erythro-) alpha,beta-epoxy alcohols in high yields with high degree of chemo- and stereoselectivity. Copyright (C) 1996 Elsevier Science Ltd

Structure of 4,6-Diacetylresorcinol

C10H10O4, M(r) = 194.19, monoclinic, P2(1)/c, a = 7.089 (1), b = 11.361 (1), c = 11.656 (1) angstrom, beta = 100.45 (3)-degrees, V = 922.92 (1) angstrom 3, Z = 4, D(m) = 1.410 (5), D(x) = 1.397 Mg m-3, lambda(Cu K-alpha) = 1.5418 angstrom, mu(Cu K-alpha) = 0.89 mm-1, T = 300 K, F(000) = 408, final R = 0.057 for 1701 observed reflections. The molecule is almost planar, with O(9) and O(12) of the acetyl groups deviating by 0.074 (1) and 0.071 (2) angstrom from the mean plane of the benzene ring. The bond lengths and bond angles of the benzene ring are normal. There are intramolecular hydrogen bonds between O(9) and H(14) and between O(12) and H(13); there are no intermolecular hydrogen bonds. The molecules are packed in layers parallel to the ac plane and are held together essentially by van der Waals interactions.

Structure of 7-ethylamino-6-methyl-4-trifluoromethylcoumarin

C13H12F3NO2, M(r) = 271.2, triclinic, P1BAR, a = 5.029 (2), b = 7.479 (2), c = 17.073 (5) angstrom, alpha = 97.98 (2), beta = 95.54 (3), gamma = 103.62 (3)-degrees, V = 612.4 (4) angstrom 3, Z = 2, D(m) = 1.463, D(x) = 1.471 g cm-3, lambda(Mo K-alpha) = 0.71069 angstrom, mu = 1.23 cm-1, F(000) = 280, T = 298 K, final R value is 0.041 for 2047 observed reflections with \F(omicron)\ greater-than-or-equal-to 6-sigma(\F(omicron)\). The N-C(sp2) bond length is 1.356 (2) angstrom. The N and C atoms of the ethylamino group deviate by < 0.15 angstrom from the plane of the aromatic ring. Short intramolecular contacts, C(3)...F(17) 2.668 (3) angstrom [H(3)...F(17) 2.39 (2) angstrom, C(3)-H(C3)...F(17) 98 (1)-degrees], C(5)...F(18) 3.074 (3) and C(5)...F(19) 3.077 (3) angstrom exist in the structure. The crystal structure is stabilized by intermolecular N-H...O hydrogen bonds with N(12)-H(N12) 0.79 (3), H(N12)...O(11)' 2.36 (3), N(12)...O(11)' (x - 1, y + 1, z) 3.105 (3) angstrom and N(12)-H(N12)...O(11)' 155 (2)-degrees.

The effect of malathion, an organophosphate, on the plasma FSH, 17 beta-estradiol and progesterone concentrations and acetylcholinesterase activity and conception in dairy cattle

The effect of malathion on jugular plasma concentrations of follicle-stimulating hormone (FSH), estradiol (E2), progesterone (P4) and acetylcholinesterase (AchE) on conception in dairy cattle during a cloprostenol (prostaglandin F2-alpha analogue, PG)-induced estrus was studied. Malathion (1 mg/kg, intraruminally) given at the onset of estrus (48 h after PG) did not alter the plasma FSH or E2 concentrations but significantly (P < 0.05) inhibited plasma P4 concentration. The mean P4 concentration in the malathion-treated group on days 8 and 12 were 0.8 +/- 0.4 and 1.0 +/- 0.5 ng/ml, as compared to 2.6 +/- 0.0 and 2.4 +/- 0.3 ng/ml in the control group. There was a nonsignificant (P > 0.05) inhibition of plasma AchE activity in malathion-treated cattle. Conception was 16.6% in malathion-treated cows and 50% in controls. Inhibition of progesterone secretion and poor conception occurred after the single intraruminal dose of malathion at the onset of estrus.