Conformation of the flexible bent helix of Leu1-zervamicin in crystal C and a possible gating action for ion passage

The membrane channel-forming polypeptide, Leu(1)-zervamicin, Ac-Leu-Ile-Gln-Iva-Ile(5)-Thr-Aib-Leu-Aib-Hyp(10) -Gln-Aib-Hyp-Aib-Pro(15)-Phol (Aib: alpha-aminoisobutyric acid; Iva: isovaline; Hyp: 4-hydroxyproline; Phol: phenylalininol) has been analyzed by x-ray diffraction in a third crystal form. Although the bent helix is quite similar to the conformations found in crystals A and B, the amount of bending is more severe with a bending angle approximate to 47 degrees, The water channel formed by the convex polar faces of neighboring helices is larger at the mouth than in crystals A and B, and the water sites have become disordered. The channel is interrupted in the middle by a hydrogen bond between the OH of Hyp(10) and the NH2 of the Gln(11) of a neighboring molecule. The side chain of Gln(11) is wrapped around the helix backbone in an unusual fashion in order that it can augment the polar side of the helix. In the present crystal C there appears to be an additional conformation for the Gln(11) side chain (with approximate to 20% occupancy) that opens the channel for possible ion passage. Structure parameters for C85H140N18O22.xH(2)O.C2H5OH are space group P2(1)2(1)2(1), a = 10.337 (2) Angstrom, b = 28.387 (7) Angstrom, c = 39.864 (11) Angstrom, Z = 4, agreement factor R = 12.99% for 3250 data observed > 3 sigma(F), resolution = 1.2 Angstrom. (C) 1994 John Wiley & Sons, Inc.

Structural plasticity and enzyme action: Crystal structures of Mycobacterium tuberculosis peptidyl-tRNA hydrolase

Peptidyl-tRNA hydrolase cleaves the ester bond between tRNA and the attached peptide in peptidyl-tRNA in order to avoid the toxicity resulting from its accumulation and to free the tRNA available for further rounds in protein synthesis. The structure of the enzyme from Mycobacteritan tuberculosis has been determined in three crystal forms. This structure and the structure of the enzyme frorn Escherichia coli in its crystal differ substantially on account of the binding of the C terminus of the E. coli enzyme to the peptide-binding site of a neighboring molecule in the crystal. A detailed examination of this difference led to an elucidation of the plasticity of the binding site of the enzyme. The peptide-binding site of the enzyme is a cleft between the body, of the molecule and a polypepticle Y stretch involving a loop and a helix. This stretch is in the open conformation when the enzyme is in the free state as in the crystals of M. tuberculosis peptidyl-tRNA hydrolase. Furthermore, there is no physical continuity between the tRNA and the peptide-binding sites. The molecule in the E. coli crystal mimics the peptide-bound enzyme molecule. The peptide stretch referred to earlier now closes on the bound peptide. Concurrently, a channel connecting the tRNA and the peptide-binding site opens primarily through the concerted movement of two residues. Thus, the crystal structure of M. tuberculosis peptidyl-tRNA hydrolase when compared with the crystal structure of the E. coli enzyme, leads to a model of structural changes associated with enzyme action on the basis of the plasticity of the molecule. (c) 2007 Elsevier Ltd. All rights reserved.

Action of N-bromosuccinimide on sperm whale myoglobin

Previously, it was reported from this laboratory that the heme groups of hemoglobin are “buried” within globin at pH 4.0 and not dissociated, on the basis of the obiligatory requirement of urea for the reaction of N-bromosuccinimide with the heme groups of hemoglobin at pH4.0, and also on the basis of the “normalization” of the spectrum of hemoglobin at this pH in the presence of urea or sucrose. In the present study, it has been shown that the behaviour of sperm whale myoglobin with respect to its reaction with N-bromosuccinimide and with respect to spectral “normalization” in urea or sucrose are essentially similar to that of hemoglobin. It has also been demonstrated that the spectral “normalization” obtained with crystalline hemin is not identical with that obtained with either hemoglobin or myoglobin. The bearing of the results of the present study on the earlier work on hemoglobin is indicated.

Action of N-bromosuccinimide on human hemoglobin and its possible bearing on heme-globin linkage

A reaction of N-bromosuccinimide with the heme groups of hemoglobin has been studied spectrophotometrically. The reaction brings about the disappearance of characteristic absorption peaks of hemoglobin and is accompanied by the release of inorganic iron from the heme groups. Urea is obligatory for the reaction to take place at pH 4.0, while it can occur in the absence of urea at pH 7.0. The spectrum of hemoglobin which does not show any peak in the Soret region at pH 4.0 is “normalized” in the presence of urea or sucrose at the same pH. The effect of “normalization” in 8 M urea is apparent over the pH range 3.0–4.5. From the obligatory requirement of urea and sucrose for “normalization” of spectrum and the dependence of the release of inorganic iron on the concentration of urea, it is suggested that heme groups are “buried” within the globin at pH 4.0 and not dissociated from globin as supposed before.

Studies in sesquiterpenes-XIX. Action of perbenzoic acid on longifolene

Oxidation of longifolene, a mono-olefinic sesquiterpene, with a chloroform solution of perbenzoic acid proceeded to almost two mole consumption of the peracid to furnish a number of products, important ones being an α-ketol, a norketone (longicamphenilone) and the corresponding C14-alcohols. Under certain conditions it has been possible to arrest the reaction at one mole consumption and to isolate longifolene-α-epoxide, which is the major product; some amounts of epimeric longifolaldehydes and longicamphenilone were also formed. Further action of perbenzoic acid on longifolene oxide and longifolaldehydes has been investigated and the results used to interpret the mechanism of abnormal peracid oxidation of longifolene.

Indoleacetaldoxime hydro-lyase (4.2.1.29). III. Further studies on the nature and mode of action of the enzyme

Further purification of indoleacetaldoxime (IAOX) hydro-lyase from Gibberella fujikuroi by DEAE-cellulose chromatography is described. The purified enzyme was activated by dehydroascorbic acid (DHA), ascorbic acid (AA), and pyridoxal phosphate (PALP) and was inhibited by thiol compounds and thiol reagents including phenylthiocyanate. Ferrous ions but not ferric ions activated the purified enzyme. The enzyme was activated by dihydrofolic acid but inhibited by tetrahydrofolic acid. Phenylacetaldoxime, a competitive inhibitor, afforded partial protection of the enzyme from the action of N-ethylmaleimide suggesting the involvement of a thiol function at the active site or substrate-binding site. The inhibition of the enzyme by 2,3-dimercaptopropanol was reversed by DHA, PALP, or frozen storage. KCN inhibition of the enzyme was reversed by PALP. NaBH4 reduction of the purified enzyme in the presence of PALP gave an active enzyme which was further activated by PALP or DHA but not by ferrous ions. These results suggested a "structural" role for PALP in the activity of IAOX hydro-lyase. Dilute solutions of the purified enzyme, obtained during DEAE-cellulose chromatography and concentrated using sucrose, showed enhanced activity upon frozen storage and thawing. The increase in activity of the enzyme during certain culture conditions, the activation and inhibition of the enzyme by several unrelated compounds, and the effect of freezing indicate that IAOX hydro-lyase is probably a metabolically regulated enzyme with a structure composed of subunits.

Ultimate Strength of RC Wall Panels in One-Way In-Plane Action

Test results of 24 reinforced concrete wall panels in one-way in-plane action are presented. The panels were loaded at a small eccentricity to reflect possible eccentric loading in practice. Influences of slenderness ratio, aspect ratio, vertical steel, and horizontal steel on the ultimate load are studied. An empirical equation modifying two existing methods is proposed for the prediction of ultimate load. The modified equation includes the effects of slenderness ratio, amount of vertical steel, and aspect ratio. The results predicted by the proposed modified method and five other available equations are compared with 48 test data. The proposed modified equation is found to be satisfactory and, additionally, includes the effect of aspect ratio which is not present in other methods.

Coding for Two-User Gaussian MAC with PSK and PAM Signal Sets

Constellation Constrained (CC) capacity regions of a two-user Gaussian Multiple Access Channel(GMAC) have been recently reported. For such a channel, code pairs based on trellis coded modulation are proposed in this paper with MPSK and M-PAM alphabet pairs, for arbitrary values of M,toachieve sum rates close to the CC sum capacity of the GMAC. In particular, the structure of the sum alphabets of M-PSK and M-PAMmalphabet pairs are exploited to prove that, for certain angles of rotation between the alphabets, Ungerboeck labelling on the trellis of each user maximizes the guaranteed squared Euclidean distance of the sum trellis. Hence, such a labelling scheme can be used systematically,to construct trellis code pairs to achieve sum rates close to the CC sum capacity. More importantly, it is shown for the first time that ML decoding complexity at the destination is significantly reduced when M-PAM alphabet pairs are employed with almost no loss in the sum capacity.

Mode of action of antitumour antibiotics-III: Modulation of permeability of nuclear membrane in the presence of the antibiotics

The binding characteristics of the antibiotics to nuclei and their effect on the permeability of nuclear membrane with respect to histones and ribonucleic acids have been investigated. The binding constant for chromomycin A3 was found to be 1.4 × 104M?1 and number of binding sites was equal to 3.48 ± 1.08 × 1012 molecules/nuclei. The antibiotic chromomycin A3 enhanced the uptake of lysine-rich histone, actinomycin D decreased the uptake and ethidium bromide had no effect. Chromomycin A3 also enhanced the release of acid insoluble fraction containing RNA from the nuclei, actinomycin D and ethidium bromide inhibited the release of acid insoluble fraction containing RNA. The relevance of this finding to the role of nuclear envelope in understanding the mechanism of action of the antibiotic has been discussed.

Comparative measurements of HV impulses to evaluate different sets of response parameters

Results are reported of comparative measurements made in 14 HV (high-voltage) laboratories in ten different countries. The theory of the proposed methods of characterizing the dynamic behavior is given, and the parameters to be used are discussed. Comparative measurements made using 95 systems based on 53 dividers are analyzed. This analysis shows that many of the system now in use, even though they fulfil the basic response requirements of the standards, do not meet the accuracy requirements. Because no transfer measurements were made between laboratories, there is no way to detect similar errors in both the system under test and the reference system. Hence, the situation may be worse than reported. This has led to the recommendation that comparative measurements should be the main route for quantifying industrial impulse measuring systems