ABSTRACT
Vicilins (7S storage proteins) found in various legume seeds have been previously shown to interfere with the germination of spores or conidia of phytopathogenic fungi and inhibit yeast growth and glucose stimulated acidification of the medium by yeast cells. In the present work vicilins from cowpea (Vigna unguiculata) seeds were added to the growth medium of Saccharomyces cerevisiae cells and Fusarium oxysporum conidia. Helix pomatia lectin, wheat germ agglutinin and Ulex europaeus lectin were used to identify differences in the binding of the vicilins to the surface of cells of S. cerevisiae and F. oxysporum treated with this protein. After the growth period, the material in suspension (yeast cells) was centrifuged and the final pellet was also treated with different sugar (glucose, sucrose, glucosamine, N-acetyl-glucosamine) concentrations and 0.1 M HCl for extraction of vicilins associated to chitinous structures present in yeast cells. Our results showed that vicilin sub-units were present in the different sugar extracts of yeast cells pretreated with the vicilins and these proteins were eluted by 0.5 M solutions of sugars in the following order of efficiency of elution: N-acetyl-glucosamine, sucrose/glucose and glucosamine.
Subject(s)
Carbohydrates/pharmacology , Binding, Competitive/drug effects , Cell Membrane/drug effects , Cell Wall/drug effects , Plant Proteins/pharmacology , Acetylglucosamine/pharmacology , Fungi/drug effects , Fungi/growth & development , Fungi/ultrastructure , Fusarium/drug effects , Fusarium/growth & development , Fusarium/ultrastructure , Glucosamine/pharmacology , Glucose/pharmacology , Binding, Competitive/physiology , Microscopy, Electron , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cell Wall/metabolism , Cell Wall/ultrastructure , Sucrose/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae/ultrastructure , Binding Sites/drug effects , Binding Sites/physiologyABSTRACT
Los receptores muscarínicos pre- y post sinápticos del vas deferens de la rata no son M1 ya que el antagonista muscarínico M1-selectivo pirenzepina (PZ) posee baja afinidad por ambos. Basándonos en este hecho las dos acciones de ACh, pre-0y post-sinápticas, en esta preparación parecen ser mediadas por receptores muscarínicos parecidos al subtipo M2. La siguiente serie de observaciones experimentales revelan que ambas respuestas son mediadas por receptores muscarínicos farmacológicamente distintos. El rango de orden de potencia desplegado por 3 antagonistas muscarínicos (Atropina, N-metil-escopolamina [NMS] y PZ) en cada uno de estos lugares son diferentes. Atropina y PZ son bloqueadores selectivos del receptor muscarínico presente en músculo liso. NMS es un antagonista selectivo del receptor pre-sináptico muscarínico que facilita la liberación de norepinefrina. Por último, PZ y NMS despliegan un antagonismo diferencial, siendo competitivos y no-competitivos pre- y post-sinápticamente, respectivamente. Los resultados sugieren que el receptor muscarínico post-sináptico presente en el músculo liso pertenece a los subtipos M2B (oM3). El receptor pre-sináptico pertenece a los subtipos M2A (o M2) o a una subclase de los receptores muscarínicos M2B (o M3)