{"id":6518,"date":"2023-02-17T07:11:27","date_gmt":"2023-02-17T07:11:27","guid":{"rendered":"https:\/\/beta.nanion.de\/resource_library\/2021-functional-characterization-of-slc-transporters-using-solid-supported-membranes\/"},"modified":"2023-02-17T07:11:27","modified_gmt":"2023-02-17T07:11:27","slug":"2021-functional-characterization-of-slc-transporters-using-solid-supported-membranes","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/ja\/resource_library\/2021-functional-characterization-of-slc-transporters-using-solid-supported-membranes\/","title":{"rendered":"2021 &#8211; Functional Characterization of SLC Transporters Using Solid Supported Membranes"},"content":{"rendered":"<p>Here, we present a protocol for the functional characterization of the H+-coupled human peptide transporter PepT1 and sufficient notes to transfer the protocol to the Na<sup>+<\/sup>-coupled sugar transporter SGLT1, the organic cation transporter OCT2, the Na<sup>+<\/sup>\/Ca<sup>2+<\/sup>\u00a0exchanger NCX, and the neuronal glutamate transporter EAAT3.The assay was developed for the commercially available SURFE2R N1 instrument (Nanion Technologies GmbH) which applies solid supported membrane (SSM)-based electrophysiology. This technique is widely used for the functional characterization of membrane transporters with more than 100 different transporters characterized so far. The technique is cost-effective, easy to use, and capable of high-throughput measurements.SSM-based electrophysiology utilizes SSM-coated gold sensors to physically adsorb membrane vesicles containing the protein of interest. A fast solution exchange provides the substrate and activates transport. For the measurement of PepT1 activity, we applied a peptide concentration jump to activate H+\/peptide symport. Proton influx charges the sensor. A capacitive current is measured reflecting the transport activity of PepT1. Multiple measurements on the same sensor allow for comparison of transport activity under different conditions. Here, we determine EC<sub>50<\/sub>\u00a0for PepT1-mediated glycylglycine transport and perform an inhibition experiment using the specific peptide inhibitor Lys[Z(NO2)]-Val.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[36],"resource_type":[75],"application_areas":[108],"therapeutic_areas":[],"target_categories":[60],"target_families":[374,375],"targets":[133,240],"resource_keyword":[],"class_list":["post-6518","resource_library","type-resource_library","status-publish","hentry","products_tags-surfe2r-n1","resource_type-publication","application_areas-transporters","target_categories-transporters","target_families-uniporter","target_families-symporter","targets-eaat3-slc1a1","targets-oct2-slc22a2"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library\/6518","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/media?parent=6518"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/products_tags?post=6518"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_type?post=6518"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/application_areas?post=6518"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/therapeutic_areas?post=6518"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_categories?post=6518"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_families?post=6518"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/targets?post=6518"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_keyword?post=6518"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}