INVESTIGADORES
COLMAN LERNER Alejandro Ariel
congresos y reuniones científicas
Título:
MEASUREMENT OF EFFECTIVE SCAFFOLD PROTEIN AFFINITY TO MEMBRANE BINDING SITES BY IN VIVO SCATCHARD
Autor/es:
ALAN BUSH; CHERNOMORETZ A; ALEJANDRO COLMAN LERNER
Lugar:
Puerto Madryn - Chubut
Reunión:
Congreso; SAIB 46th Annual Meeting Argentine Society for Biochemistry and Molecular Biology XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular.; 2010
Institución organizadora:
Society for Biochemistry and Molecular Biology XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular.
Resumen:
The yeast, Saccharomyces cerevisiae, pheromone response
pathway is one of the best understood signal transduction systems.
All key components of the system and their interactions have been
identified by more than 30 years of genetic and biochemical
investigation. In recent years modeling efforts have been
undertaken to try to describe the behavior of the system in a
quantitative manner. These models usually contain a large number
of parameters to be determined by fitting to a dataset of
experimental results. Direct measurement of these parameters can
be used to constrain these models and obtain more accurate and
predictive descriptions of the pathway.
In this work we describe a microscopy imaging technique
developed to measure protein relocalization in yeast in a
quantitative manner. Using this technique we measured membrane
recruitment of the scaffold protein of the mating pathway Ste5, an
early event that occurs upon pheromone stimulation. Varying the
amount of Ste5 by genetic manipulation allowed us to perform anSaccharomyces cerevisiae, pheromone response
pathway is one of the best understood signal transduction systems.
All key components of the system and their interactions have been
identified by more than 30 years of genetic and biochemical
investigation. In recent years modeling efforts have been
undertaken to try to describe the behavior of the system in a
quantitative manner. These models usually contain a large number
of parameters to be determined by fitting to a dataset of
experimental results. Direct measurement of these parameters can
be used to constrain these models and obtain more accurate and
predictive descriptions of the pathway.
In this work we describe a microscopy imaging technique
developed to measure protein relocalization in yeast in a
quantitative manner. Using this technique we measured membrane
recruitment of the scaffold protein of the mating pathway Ste5, an
early event that occurs upon pheromone stimulation. Varying the
amount of Ste5 by genetic manipulation allowed us to perform an
in vivo Scatchard to measure the effective binding affinity of the
scaffold protein with its membrane associated binding sites. Using
this parameter we will refine our mathematical model of the onset
of the pheromone response pathway.Scatchard to measure the effective binding affinity of the
scaffold protein with its membrane associated binding sites. Using
this parameter we will refine our mathematical model of the onset
of the pheromone response pathway.