Home Abstract Introduction
Construction of glutamate sensors Initial testing of sensors in vitro Optimization of sensor characteristics Expression of sensor in living cells Glutamate measurement in dendrites Glutamate measurement in response to frequency of action potentials
Glutamate function Sensor design
Protein purification through His tags Overexpression of protein using pRSETB Directing proteins to a membrane by signal peptides Transmembrane regions of proteins Expression of proteins on the surface of mammalian cells through pDISPLAY

Journal of Comprehensible Results

Hires SA, Zhu Y, Tsien R (2008)
Optical measurement of synaptic glutamate spillover and reuptake
by linker optimized glutamate-sensitive fluorescent reporters
Proc Natl Acad Sci USA 105:4411-4416

(Translated by Jeff Elhai)

Implications: Sensor design

[Two means of optimization: mutagenesis of glutamate binding protein to reduce affinity and truncation of the protein to increase +/- glutamate difference.

Optimized sensor far better than original, but difficult to predict best way to truncate the glutamate binding protein to achieve optimization.]