• Refine Query
  • Source
  • Publication year
  • to
  • Language
  • 1
  • Tagged with
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Papel de proteÃnas presentes na tinta liberada pela lesma-do-mar Aplysia dactylomela Rang 1828 nos mecanismos de defesa do animal / Role of protein present in the ink released by-the-sea slug Aplysia dactylomela Rang in 1828 in defense mechanisms of the animal

Vanessa Lucia Rodrigues Nogueira 07 December 2005 (has links)
O gastrÃpode marinho Aplysia dactylomela à conhecido por liberar uma tinta pÃrpura sempre que à importunado. Como nÃo possui nenhuma estrutura externa de proteÃÃo, acredita-se que essa secreÃÃo, rica em substÃncias biologicamente ativas, participe da defesa quÃmica do animal. A tinta à composta de pigmentos, proteÃnas e substÃncias de baixa massa molecular. Os pigmentos da tinta sÃo originados de algas vermelhas, mas quanto Ãs proteÃnas, pouco à conhecido sobre a sua origem, processamento, local de armazenamento e funÃÃo no animal. Este trabalho descreve pela primeira vez a composiÃÃo protÃica da tinta e apresenta algumas caracterÃsticas fÃsico-quÃmicas e biolÃgicas da dactylomelina-P, uma proteÃna antibacteriana presente na tinta dessa lesma, alÃm de trazer informaÃÃes acerca da localizaÃÃo desta proteÃna na glÃndula de tinta. A tinta foi obtida a partir de espÃcimes encontrados na praia de Fleixeiras, Ce. A composiÃÃo de proteÃnas da tinta foi determinada por eletroforese bidimensional e a purificaÃÃo da dactylomelina-P foi feita atravÃs de cromatografias de troca iÃnica e interaÃÃo hidrofÃbica. Dactylomelina-P foi analisada quanto à massa molecular, ponto isoelÃtrico, composiÃÃo de aminoÃcidos, presenÃa de carboidratos, estabilidade tÃrmica, resistÃncia ao pH e a proteases. VÃrias atividades biolÃgicas foram testadas com a tinta e com a dactylomelina-P, incluindo atividades antimicrobianas, enzimÃticas, hemaglutinante, anticoagulante, hemolÃtica, citotÃxica e tÃxica. Os ensaios de localizaÃÃo da proteÃna foram realizados em diferentes tecidos da lesma por western blot na glÃndula de tinta por imunohistoquÃmica e a interaÃÃo com a bactÃria Staphylococcus foi feita por imunocitoquÃmica. A tinta de A. dactylomela contÃm mais de 40 proteÃnas/ peptÃdeos, com massas abaixo de 70 kDa e pIs na faixa Ãcida. A proteÃna mais abundante na tinta à a dactylomelina-P, que à uma molÃcula monomÃrica, de 59,8 kDa, pI 5,0, que possui alto teor de metionina e menos de 1% de carboidratos, à desnaturada a 60 ÂC por 10 minutos e resiste a pHs entre 3-12. Dactylomelina-P mostra um amplo espectro de aÃÃo antibacteriano, mas nÃo antifÃngico, ao contrÃrio da tinta que possui um fator com esta atividade. à particularmente eficiente contra bactÃrias marinhas, podendo ser bactericida (4,0μg/ml) ou bacteriostÃtica (0,2μg/ml), dependendo da concentraÃÃo. Dactylomelina-P aglutinou eritrÃcitos de coelho, ratos e camundongos, nÃo apresentou atividade anticoagulante, hemolÃtica e nem citotÃxica. A DL50 para camundongos ficou entre 60-100 mg/Kg, sendo considerada moderadamente tÃxica. Dactylomelina-P sà foi encontrada na glÃndula de tinta, localizando-se preferencialmente nas cÃlulas das vesÃculas produtoras. Ensaios de microscopia eletrÃnica de transmissÃo revelaram que a dactylomelina atravessa a parede celular da bactÃria S. aureus e, interage principalmente com a membrana citoplasmÃtica, provavelmente interferindo no metabolismo, ao invÃs de causar danos à cÃlula. / The marine gastropod Aplysia dactylomela is known to release a purple ink when it is plagued. How has no external structure of protection, it is believed that secretion, rich in biologically active substances, participate in the chemical defense of the animal. The ink is composed of pigments, proteins and low molecular weight substances. The pigments of ink are originated from red algae, but as the protein, little is known about its origin, processing, storage location and function in animals. This paper describes for the first time the protein composition of the ink and presents some physico-chemical and dactylomelina of organic-P, an antibacterial protein present in the ink that slug, and get information about the location of this protein in the ink gland. The paint was obtained from specimens found on the beach in Fleixeiras, Ce. The protein composition of the ink was determined by two-dimensional electrophoresis and purification of dactylomelina-P was done by ion exchange chromatography and hydrophobic interaction. Dactylomelina-P was analyzed on the molecular mass, isoelectric point, composition of amino acids, the presence of carbohydrates, thermal stability, resistance to proteases and pH. Several biological activities were tested with the ink and the dactylomelina-P, including antimicrobial activity, enzyme, haemagglutinating, anticoagulant, hemolytic, cytotoxic and toxic. Tests for localization of the protein were performed in different tissues of the slug by western blot in the ink gland by immunohistochemistry and the interaction with the bacterium Staphylococcus was performed by immunocytochemistry. The ink of A. dactylomela contains more than 40 proteins / peptides, with masses below 70 kDa and PIs in the acidic range. The most abundant protein in the ink is dactylomelina-P, which is a monomeric molecule of 59.8 kDa, pI 5.0, which has high content of methionine and less than 1% of carbohydrates, is denatured at 60  C for 10 minutes and resist the pH between 3-12. Dactylomelina-P shows a broad spectrum of antibacterial action, but not antifungal, unlike paint that has a factor with this activity. It is particularly effective against marine bacteria and may be bactericidal (4.0 g / ml) or bacteriostatic (0.2 g / ml), depending on the concentration. P-Dactylomelina agglutinate rabbit erythrocytes, rats and mice, showed no anticoagulant activity, and cytotoxic and hemolytic. The LD50 for mice was between 60-100 mg / kg, is considered moderately toxic. Dactylomelina-P was found in the ink gland, is located preferentially in cells producing the vesicles. Tests for transmission electron microscopy revealed that the dactylomelina through the cell wall of the bacterium S. aureus and mainly interacts with the cytoplasmic membrane, probably interfering with metabolism, rather than damage the cell.

Page generated in 0.344 seconds