{"id":4715,"date":"2023-02-17T07:09:43","date_gmt":"2023-02-17T07:09:43","guid":{"rendered":"http:\/\/ionchanneltargets.com\/resource_library\/acetylcholine-receptor-alpha-3-beta-4-nicotinic-a3b4-receptors-recorded-on-nanions-patchliner\/"},"modified":"2023-02-17T07:09:44","modified_gmt":"2023-02-17T07:09:44","slug":"2006-structure-and-function-of-prokaryotic-glutamate-transporters-from-escherichia-coli-and-pyrococcus-horikoshii","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/zh-hans\/resource_library\/2006-structure-and-function-of-prokaryotic-glutamate-transporters-from-escherichia-coli-and-pyrococcus-horikoshii\/","title":{"rendered":"2006 &#8211; Structure and function of prokaryotic glutamate transporters from Escherichia coli and Pyrococcus horikoshii"},"content":{"rendered":"<p>The glutamate transporters GltPEc\u00a0from Escherichia coli and GltPPh\u00a0from Pyrococcus horikoshii were overexpressed in E. coli and purified to homogeneity with a yield of 1-2 mg\/L of culture. Single-particle analysis and electron microscopy indicate that GltP(Ph) is a trimer in detergent solution. Electron microscopy of negatively stained GltPPh\u00a0two-dimensional crystals shows that the transporter is a trimer also in the membrane. Gel filtration of GltPEc\u00a0indicates a reversible equilibrium of two oligomeric states in detergent solution that we identified as a trimer and hexamer by blue-native gel electrophoresis and cross-linking. The purified transporters were fully active upon reconstitution into liposomes, as demonstrated by the uptake of radioactively labeled L-aspartate or L-glutamate. L-aspartate\/L-glutamate transport of GltPEc\u00a0involves the cotransport of protons and depends only on pH, whereas GltP(Ph) catalyzes L-glutamate transport with a cotransport of H+\u00a0or Na<sup>+<\/sup>. L-glutamate induces a fast transient current in GltP(Ph) proteoliposomes coupled to a solid supported membrane (SSM). We show that the electric signal depends on the concentration of Na<sup>+<\/sup>\u00a0or H+\u00a0outside the proteoliposomes and that GltP(Ph) does not require K<sup>+<\/sup>\u00a0inside the proteoliposomes. In addition, the electrical currents are inhibited by TBOA and HIP-B. The half-saturation concentration for activation of GltPPh\u00a0glutamate transport (K0.5glut) is 194 \u00b5M.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[36],"resource_type":[75],"application_areas":[108],"therapeutic_areas":[],"target_categories":[60],"target_families":[],"targets":[],"resource_keyword":[],"class_list":["post-4715","resource_library","type-resource_library","status-publish","hentry","products_tags-surfe2r-n1","resource_type-publication","application_areas-transporters","target_categories-transporters"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library\/4715","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/media?parent=4715"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/products_tags?post=4715"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_type?post=4715"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/application_areas?post=4715"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/therapeutic_areas?post=4715"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_categories?post=4715"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_families?post=4715"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/targets?post=4715"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_keyword?post=4715"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}