Differential sensitivity of stilbenedisulfonates in their reaction with band 3 HT (Pro-868-->Leu).

Band 3 HT (Pro-868-->Leu) is a mutant anion exchange protein which has several phenotypic characteristics, including a 2- to 3-fold larger Vmax, and reduced covalent binding of the anion transport inhibitor 4,4'-diisothiocyanodihydrostilbene-2,2'-disulfonate (H2DIDS). We have used fluorescence kinetic methods to study inhibitor binding to band 3 to determine if the point mutation in band 3 HT produces localized or wide-spread conformational changes within the membrane-bound domain of this transporter. Our results show that covalent binding of H2DIDS by band 3 HT is slower by a factor of 10 to 20 compared with the wild-type protein. In contrast, no such difference in the kinetics was observed for covalent binding of 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS). In addition, the kinetics of H2DIDS release from band 3 HT was abnormal, while the kinetics of 4,4'-dibenzamidostilbene-2,2'-disulfonate (DBDS) release showed no difference when compared with the wild-type protein. We conclude that substitution of leucine for proline at position 868 does not perturb the structure of "lysine A" in the membrane-bound domain of band 3 but rather produces an apparently localized conformational change in the C-terminal subdomain of the protein which alters H2DIDS affinity. When combined with the observation of an increased Vmax, these results suggest that protein structural changes at position 868 influence a turnover step in the transport cycle.

Role of the integument in insect defense: pro-phenol oxidase cascade in the cuticular matrix.

The cuticle of the silkworm Bombyx mori was demonstrated to contain pro-phenol oxidase [zymogen of phenol oxidase (monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1)] and its activating cascade. The activating cascade contained at least one serine proteinase zymogen (latent form of pro-phenol oxidase activating enzyme). When the extracted cascade components were incubated with Ca2+, the latent form of pro-phenol oxidase activating enzyme was itself activated and, in turn, converted through a limited proteolysis of pro-phenol oxidase to phenol oxidase. Immuno-gold localization of prophenol oxidase in the cuticle using a cross-reactive hemolymph anti-pro-phenol oxidase antibody revealed a random distribution of this enzyme in the nonlamellate endocuticle and a specific orderly arrayed pattern along the basal border of the laminae in the lamellate endocuticle of the body wall. Furthermore, prophenol oxidase was randomly distributed in the taenidial cushion of the tracheal cuticle. At the time of pro-phenol oxidase accumulation in the body wall cuticle, no pro-phenol oxidase mRNA could be detected in the epidermal tissue, whereas free-circulating hemocytes contained numerous transcripts of pro-phenol oxidase. Our results suggest that the pro-phenol oxidase is synthesized in the hemocytes and actively transported into the cuticle via the epidermis.

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.

Índice de BICs del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante)

Índice de BICs del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante), sin aprobación definitiva.

Ficha (1ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante)

Ficha (1ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante), sin aprobación definitiva.

Ficha (2ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante)

Ficha (2ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante), sin aprobación definitiva.

Ficha (3ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante)

Ficha (3ª) del Faro del Cabo de Huertas del PEPTHA (Plan Especial de Protección de las Torres de la Huerta de Alicante), sin aprobación definitiva.

Reflexiones sobre la huerta y propuestas para su conservación. VII Jornadas en Defensa de la Huerta (La huerta, código genético del territorio y la sociedad)

Cátedra Arzobispo Loazes, Universidad de Alicante.

Cambios históricos en el patrón del paisaje agrícola en un sistema de huerta tradicional de la provincia de Alicante: el caso de la Finca Lo Comptador

Este estudio de caso de un paisaje agrícola en un sistema de huerta tradicional de la provincia de Alicante fue llevado a cabo para desarrollar técnicas adecuadas de cuantificación y análisis del paisaje en el período 1883-2007. Los métodos utilizados se derivan del análisis de cambio en la estructura del paisaje, basado en antiguos mapas y fotografías aéreas, siendo adaptados a una escala detallada por medio de herramientas SIG. La base de datos creada para este análisis diacrónico es una pieza fundamental para obtener una clara visión de la evolución de los usos del suelo en el área de estudio. El principal objetivo de este estudio es poner de manifiesto que una aproximación a nivel de parcela, utilizando tecnologías SIG, proporciona valiosos resultados para planificar programas de conservación en paisajes agrícolas a escala local.