Molecular cloning of insect pro-phenol oxidase: a copper-containing protein homologous to arthropod hemocyanin.

Pro-phenol oxidase [pro-PO; zymogen of phenol oxidase (monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1)] is present in the hemolymph plasma of the silkworm Bombyx mori. Pro-PO is a heterodimeric protein synthesized by hemocytes. A specific serine proteinase activates both subunits through a limited proteolysis. The amino acid sequences of both subunits were deduced from their respective cDNAs; amino acid sequence homology between the subunits was 51%. The deduced amino acid sequences revealed domains highly homologous to the copper-binding site sequences (copper-binding sites A and B) of arthropod hemocyanins. The overall sequence homology between silkworm pro-PO and arthropod hemocyanins ranged from 29 to 39%. Phenol oxidases from prokaryotes, fungi, and vertebrates have sequences homologous to only the copper-binding site B of arthropod hemocyanins. Thus, silkworm pro-PO DNA described here appears distinctive and more closely related to arthropod hemocyanins. The pro-PO-activating serine proteinase was shown to hydrolyze peptide bonds at the carboxyl side of arginine in the sequence-Asn-49-Arg-50-Phe-51-Gly-52- of both subunits. Amino groups of N termini of both subunits were indicated to be N-acetylated. The cDNAs of both pro-PO subunits lacked signal peptide sequences. This result supports our contention that mature pro-PO accumulates in the cytoplasm of hemocytes and is released by cell rupture, as for arthropod hemocyanins.

The effect of protein concentration on ion binding.

The concentration of protein in a solution has been found to have a significant effect on ion binding affinity. It is well known that an increase in ionic strength of the solvent medium by addition of salt modulates the ion-binding affinity of a charged protein due to electrostatic screening. In recent Monte Carlo simulations, a similar screening has been detected to arise from an increase in the concentration of the protein itself. Experimental results are presented here that verify the theoretical predictions; high concentrations of the negatively charged proteins calbindin D9k and calmodulin are found to reduce their affinity for divalent cations. The Ca(2+)-binding constant of the C-terminal site in the Asn-56 --> Ala mutant of calbindin D9k has been measured at seven different protein concentrations ranging from 27 microM to 7.35 mM by using 1H NMR. A 94% reduction in affinity is observed when going from the lowest to the highest protein concentration. For calmodulin, we have measured the average Mg(2+)-binding constant of sites I and II at 0.325, 1.08, and 3.25 mM protein and find a 13-fold difference between the two extremes. Monte Carlo calculations have been performed for the two cases described above to provide a direct comparison of the experimental and simulated effects of protein concentration on metal ion affinities. The overall agreement between theory and experiment is good. The results have important implications for all biological systems involving interactions between charged species.

Experimental evidence for hydrogen-bonded network proton transfer in bacteriorhodopsin shown by Fourier-transform infrared spectroscopy using azide as catalyst.

Experimental evidence for proton transfer via a hydrogen-bonded network in a membrane protein is presented. Bacteriorhodopsin's proton transfer mechanism on the proton uptake pathway between Asp-96 and the Schiff base in the M-to-N transition was determined. The slowdown of this transfer by removal of the proton donor in the Asp-96-->Asn mutant can be accelerated again by addition of small weak acid anions such as azide. Fourier-transform infrared experiments show in the Asp-96-->Asn mutant a transient protonation of azide bound to the protein in the M-to-N transition and, due to the addition of azide, restoration of the IR continuum band changes as seen in wild-type bR during proton pumping. The continuum band changes indicate fast proton transfer on the uptake pathway in a hydrogen-bonded network for wild-type bR and the Asp-96-->Asn mutant with azide. Since azide is able to catalyze proton transfer steps also in several kinetically defective bR mutants and in other membrane proteins, our finding might point to a general element of proton transfer mechanisms in proteins.

Diminished degradation of yeast cytochrome c by interactions with its physiological partners.

The level and structure of yeast iso-1-cytochrome c and iso-2-cytochrome c, encoded by the nuclear genes CYC1 and CYC7, respectively, are normally not altered in rho- mutants, which completely lack the cytochromes a.a3 subunits and cytochrome b that are encoded by mitochondrial DNA. In contrast, iso-cytochromes c containing the amino acid change Thr-78-->Ile (T78I) were observed at the normal or near-normal wild-type level in rho+ strains but were completely absent in rho- mutants. We have demonstrated with the "global" suppressor mutation Asn-52-->Ile and by pulse-chase labeling that the T78I iso-1-cytochrome c undergoes rapid cellular degradation in rho- mutants. Furthermore, specific mutations revealed that the deficiency of T78I iso-1 cytochrome c can be caused by the lack of cytochrome a.a3 or cytochrome c1, but not by the lack of cytochrome b. Thus, this and certain other, but not all, labile forms of cytochrome c are protected from degradation by the interaction with its physiological partners.

Unlocking the Value of Variation in CKD Prevalence

Peer reviewed

Recuperación de estructuras latentes y estimación de parámetros en test multidimensionales ordinales: análisis comparado entre la TRI la TCT y el análisis factorial

A través de la realización de cuatro estudios basados en datos simulados y unidos por una preocupación común, aunque parcialmente independientes en su metodología y foco específico, esta tesis se plantea como objetivo general poner a prueba algunas de las ventajas que podría obtener un investigador aplicado al utilizar modelos clásicos de TRI frente a otras opciones, para de esta forma evaluar la posibilidad de obtener resultados relativamente equivalentes utilizando procedimientos alternativos. Específicamente, en esta tesis se pone a prueba: a) la calidad diferencial de las estimaciones de las propiedades de los ítems obtenidas por los procedimientos TRI frente a los factoriales; b) la validez de los procedimientos de escalamiento TRI frente a los de la TCT; y c) los límites de la propiedad de invarianza de las estimaciones de sujetos e ítems obtenidas con procedimientos TRI. La hipótesis general de esta tesis es que, en el tipo de condiciones en que se ha enfocado el estudio, las ventajas de utilizar modelos clásicos de TRI no serán demasiado amplias, con lo que existirán condiciones en las que emplear procedimientos alternativos será recomendable, dada su simplicidad relativa y su equivalencia de resultados. La primera investigación utiliza datos simulados para comparar las estimaciones de discriminación de ítems politómicos provenientes del AFI, frente a las posibles de obtener con distintas estrategias relacionadas con el análisis factorial clásico. La segunda investigación, también utiliza datos simulados para poner a prueba la equivalencia de las estimaciones de discriminación de los ítems obtenidas con los procedimientos TRI y AFI. La tercera investigación, utiliza dos estudios basados en datos simulados tanto para comparar los escalamientos de los sujetos obtenidos por la TRI y la TCT, como para aclarar las condiciones en que es posible esperar la presencia similitudes o diferencias entre ambos...

Recensiones

Contiene: GARCÍA BALLESTER, Luis, Los Moriscos y la medicina. Un capitulo de la medicina y la ciencia marginadas en la España del siglo XVI, documentos por Rosa Blasco y Luis García Ballester, Editorial Labor, Barcelona, 1984, 256 pp. / Míkel de Epalza ; LÓPEZ ELUM, Pedro, Los orígenes de la cerámica de Manises y Paterna (1285-1335), Manises y Paterna, 1984, 99 páginas / Rafael Azuar ; ROSSELLÓ PONS, Margalida, Les ceramiques almohades del Carrer de Zavellà Ciutat de Mallorca. Palma de Mallorca, 1983, 159 páginas / Rafael Azuar ; TORNERO POVEDA, Emilio, La disputa de los animales contra el hombre (traducción del original árabe de «La disputa del asno contra Fray Anselmo Turmeda»), Madrid, Editorial de la Universidad Complutense, 1984, 234 pp. / Míkel de Epalza ; VÁZQUEZ DE BENITO, María de la Concepción, Muḥammad b. Abdallāh b. al-Jatīb. K. al-Wuṣūl li-ḥifz al-ṣiḥḥa fi-l-fuṣūl. «Libro de la Higiene», Salamanca, Ediciones Universidad de Salamanca, 1984, 282-182 páginas. - Commentaria Averrois in Galenum, Madrid, Consejo Superior de Investigaciones Científicas (Instituto «Miguel Asín») e Instituto Hispano-Árabe de Cultura, 1984, XII. 341 páginas / Míkel de Epalza.

Colossal anisotropy in diluted magnetic topological insulators

We consider dilute magnetic doping in the surface of a three dimensional topological insulator where a two dimensional Dirac electron gas resides. We find that exchange coupling between magnetic atoms and the Dirac electrons has a strong and peculiar effect on both. First, the exchange-induced single ion magnetic anisotropy is very large and favors off-plane orientation. In the case of a ferromagnetically ordered phase, we find a colossal magnetic anisotropy energy, of the order of the critical temperature. Second, a persistent electronic current circulates around the magnetic atom and, in the case of a ferromagnetic phase, around the edges of the surface.

Topological spin waves in the atomic-scale magnetic skyrmion crystal

We study the spin waves of the triangular skyrmion crystal that emerges in a two-dimensional spin lattice model as a result of the competition between Heisenberg exchange, Dzyalonshinkii–Moriya interactions, Zeeman coupling and uniaxial anisotropy. The calculated spin wave bands have a finite Berry curvature that, in some cases, leads to non-zero Chern numbers, making this system topologically distinct from conventional magnonic systems. We compute the edge spin-waves, expected from the bulk-boundary correspondence principle, and show that they are chiral, which makes them immune to elastic backscattering. Our results illustrate how topological phases can occur in self-generated emergent superlattices at the mesoscale.

Majmūʻah : manuscript, 1848-1857

1. Kitāb Maqṣad al-asná fī dhikr mā yataʻallaq bi-al-asmāʼ al-ḥusná / Aḥmad Zarrūq, 1267 AH [1850 or 51 AD] (ff. 1r-21v) -- 2. Sharḥ sayyid al-istighfār / ʻAlī Mahāyimī, 1265 AH [1848 or 49 AD] (ff. 25r-32v) -- 3. Hādhā al-suʼāl nuqila min asʼilah suʼila ʻanhā ... Muḥammad ibn Sulaymān al-Kurdī al-Shāmī al-Madanī (ff. 33r-39r) -- 4. Hādhā fawāyid nafīsah ... wa-mimmā suʼila ʻanhu ... al-Shaykh Saʻīd Sunbul al-Makkī, 1273 AH [1856 or 57 AD] (ff. 39r-51r) -- 5. Mā qawl al-sādah al-Ḥanafīyah fī ʻaṣīr qaṣab al-sukkar / ʻAbd al-Wahhāb al-Suyūṭī al-Ḥanafī, ʻAbd al-Qādir al-Rāfiʻī al-Ḥanafī, ʻAbd al-Raḥmān al-Baḥrāwī al-Ḥanafī, Muḥammad al-Rāfiʻī al-Ḥanafī (ff. 51v-55r).

Modifications post-traductionnelles des canaux calciques cardiaques de type L : identification des résidus asparagine qui participent à la glycosylation de la sous-unité auxiliaire CaVα2δ1

Les canaux calciques de type L CaV1.2 sont principalement responsables de l’entrée des ions calcium pendant la phase plateau du potentiel d’action des cardiomyocytes ventriculaires. Cet influx calcique est requis pour initier la contraction du muscle cardiaque. Le canal CaV1.2 est un complexe oligomérique qui est composé de la sous-unité principale CaVα1 et des sous-unités auxiliaires CaVβ et CaVα2δ1. CaVβ joue un rôle déterminant dans l’adressage membranaire de la sous-unité CaVα1. CaVα2δ1 stabilise l’état ouvert du canal mais le mécanisme moléculaire responsable de cette modulation n’a pas été encore identifié. Nous avons récemment montré que cette modulation requiert une expression membranaire significative de CaVα2δ1 (Bourdin et al. 2015). CaVα2δ1 est une glycoprotéine qui possède 16 sites potentiels de glycosylation de type N. Nous avons donc évalué le rôle de la glycosylation de type-N dans l’adressage membranaire et la stabilité de CaVα2δ1. Nous avons d’abord confirmé que la protéine CaVα2δ1 recombinante, telle la protéine endogène, est significativement glycosylée puisque le traitement à la PNGase F se traduit par une diminution de 50 kDa de sa masse moléculaire, ce qui est compatible avec la présence de 16 sites Asn. Il s’est avéré par ailleurs que la mutation simultanée de 6/16 sites (6xNQ) est suffisante pour 1) réduire significativement la densité de surface de! CaVα2δ1 telle que mesurée par cytométrie en flux et par imagerie confocale 2) accélérer les cinétiques de dégradation telle qu’estimée après arrêt de la synthèse protéique et 3) diminuer la modulation fonctionnelle des courants générés par CaV1.2 telle qu’évaluée par la méthode du « patch-clamp ». Les effets les plus importants ont toutefois été obtenus avec les mutants N663Q, et les doubles mutants N348Q/N468Q, N348Q/N812Q, N468Q/N812Q. Ensemble, ces résultats montrent que Asn663 et à un moindre degré Asn348, Asn468 et Asn812 contribuent à la biogenèse et la stabilité de CaVα2δ1 et confirment que la glycosylation de type N de CaVα2δ1 est nécessaire à la fonction du canal calcique cardiaque de type L.

Organic properties of sediments and amino acids in iterstitial waters of ODP Holes 111-677A and 111-678B

The basement at Ocean Drilling Program (ODP) Sites 677 and 678 originated from the Galapagos spreading center of the Costa Rica Rift and has moved about 200 km over the last 6 m.y. (Fig. 1) (Shipboard Scientific Party, 1987, 1988; Scientific Drilling Party, 1987). Sediments about 300 m thick cover basement so young that basal sediments at Sites 677 and 678 have been reheated up to 60?-70?C at Site 677 and altered to limestone and/or chert (Shipboard Scientific Party, 1988). Sediments from both sites indicate (1) a high sedimentation rate (about 48 m/m.y.) and (2) biogenic silica and carbonate as the main constituents of sediments (Table 1) (Shipboard Scientific Party, 1988). Heatflow observations and measurements of interstitial water chemistry around the sites show that Site 677 is in a lower heatflow zone (166 mW/m**2; 1°12.14'N, 83°44.22'W) whereas Site 678 is located in a zone of higher heat flow (250 mW/m**2; 1°13.01'N, 83°43.39'W) (Langseth et al., 1988; Shipboard Scientific Party, 1988). In the flank hydrothermal systems, circulating solution is moving upward through the sedimentary column in zones of higher heat flow while it is moving downward in zones of lower heat flow (Anderson and Skilbeck, 1981). The chemistry of the interstitial waters is modified by several processes such as (1) diagenetic reactions and (2) advective and (3) diffusive transports of dissolved constituents. Analyses of Ca2+ and Mg2+ in interstitial waters from Sites 677 and 678 show that their profiles are mainly controlled by advective transport (Shipboard Scientific Party, 1988). In contrast, the interstitial-water profiles for NH4+, Si, and PO4[3-] are highly affected by reactions in the sediments. Site 677 offers a good opportunity to investigate amino acids in the interstitial waters because sediments of similar compositions have been deposited at constant rates of sedimentation. There are few previous works on amino acid distributions in interstitial waters (Henrichs and Parrington, 1979; Michaelis et al., 1982; Henrichs et al., 1984; Henrichs and Farrington, 1987; Ishizuka et al., 1988). In this chapter, we report (1) Rock-Eval analysis and (2) the composition of total hydrolyzable and dissolved free amino acids (THAA and DFAA, respectively) in the interstitial waters. Our objectives are to discuss (1) the possible origin of organic materials, (2) the characteristics of THAA and DFAA, and (3) their relationships in interstitial waters.

Nachertanie statistiki rossĭskago gosudarstva /

Mode of access: Internet.

Art by the way /

One of 750 copies.