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- W2556587095 abstract "The serotonin transporter is a sodium and chloride-coupled transporter that pumps extracellular serotonin into cells. S-citalopram is a drug used to treat depression and anxiety by binding to the serotonin transporter with high-affinity, blocking serotonin reuptake. Here we report an efficient procedure and a set of tools to stabilize, express, purify, and crystallize serotonin transporter-antibody complexes bound to S-citalopram and other antidepressants. Mutations which stabilize the serotonin transporter were identified using an S-citalopram binding assay. Serotonin transporter expressed in baculovirus-transduced HEK293S GnTI- cells, was reconstituted into proteoliposomes and used to raise high-affinity antibodies. We have developed a strategy to discover antibodies that are useful for structural studies. A straightforward approach for the expression of antibody fragments in Sf9 cells has also been established. Transporter-antibody complexes purified using this procedure are well-behaved and readily crystallize, producing complexes with S-citalopram that diffract X-rays to 3-4 Å resolution. The strategies developed here can be utilized to determine the structure of other challenging membrane proteins." @default.
- W2556587095 created "2016-11-30" @default.
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- W2556587095 date "2016-11-27" @default.
- W2556587095 modified "2023-10-11" @default.
- W2556587095 title "Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to <em>S</em>-citalopram" @default.
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- W2556587095 doi "https://doi.org/10.3791/54792" @default.
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