Title : Nonoisotopic assay for the presynaptic choline transporter reveals capacity for allosteric modulation of choline uptake.

Pub. Date : 2012 Oct 17

PMID : 23077721






9 Functional Relationships(s)
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1 We established that the addition of choline to these cells, at concentrations appropriate for high-affinity choline transport at presynaptic terminals, generates a hemicholinium-3 (HC-3)-sensitive, membrane depolarization that can be used for the screening of CHT inhibitors and activators. Choline solute carrier family 5 member 7 Homo sapiens
2 We established that the addition of choline to these cells, at concentrations appropriate for high-affinity choline transport at presynaptic terminals, generates a hemicholinium-3 (HC-3)-sensitive, membrane depolarization that can be used for the screening of CHT inhibitors and activators. Choline solute carrier family 5 member 7 Homo sapiens
3 We established that the addition of choline to these cells, at concentrations appropriate for high-affinity choline transport at presynaptic terminals, generates a hemicholinium-3 (HC-3)-sensitive, membrane depolarization that can be used for the screening of CHT inhibitors and activators. Hemicholinium 3 solute carrier family 5 member 7 Homo sapiens
4 We established that the addition of choline to these cells, at concentrations appropriate for high-affinity choline transport at presynaptic terminals, generates a hemicholinium-3 (HC-3)-sensitive, membrane depolarization that can be used for the screening of CHT inhibitors and activators. Hemicholinium 3 solute carrier family 5 member 7 Homo sapiens
5 Using this assay, we discovered that staurosporine increased CHT LV-AA choline uptake activity, an effect mediated by a decrease in choline K(M) with no change in V(max). Staurosporine solute carrier family 5 member 7 Homo sapiens
6 Using this assay, we discovered that staurosporine increased CHT LV-AA choline uptake activity, an effect mediated by a decrease in choline K(M) with no change in V(max). Choline solute carrier family 5 member 7 Homo sapiens
7 Using this assay, we discovered that staurosporine increased CHT LV-AA choline uptake activity, an effect mediated by a decrease in choline K(M) with no change in V(max). Choline solute carrier family 5 member 7 Homo sapiens
8 Surprisingly, staurosporine reduced choline-induced membrane depolarization, suggesting that increased substrate coupling to ion gradients, arising at the expense of nonstoichiometric ion flow, accompanies a shift of CHT to a higher-affinity state. Staurosporine solute carrier family 5 member 7 Homo sapiens
9 Surprisingly, staurosporine reduced choline-induced membrane depolarization, suggesting that increased substrate coupling to ion gradients, arising at the expense of nonstoichiometric ion flow, accompanies a shift of CHT to a higher-affinity state. Choline solute carrier family 5 member 7 Homo sapiens