A radioligand binding assay for the insulin-like growth factor 2 receptor.
Insulin-like growth factors 2 and 1 (IGF2 and IGF1) and insulin are closely related hormones that are responsible for the regulation of metabolic homeostasis, development and growth of the organism. Physiological functions of insulin and IGF1 are relatively well-studied, but information about the ro...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0238393 |
| _version_ | 1819017109196242944 |
|---|---|
| author | Pavlo Potalitsyn Irena Selicharová Kryštof Sršeň Jelena Radosavljević Aleš Marek Kateřina Nováková Jiří Jiráček Lenka Žáková |
| author_facet | Pavlo Potalitsyn Irena Selicharová Kryštof Sršeň Jelena Radosavljević Aleš Marek Kateřina Nováková Jiří Jiráček Lenka Žáková |
| author_sort | Pavlo Potalitsyn |
| collection | DOAJ |
| description | Insulin-like growth factors 2 and 1 (IGF2 and IGF1) and insulin are closely related hormones that are responsible for the regulation of metabolic homeostasis, development and growth of the organism. Physiological functions of insulin and IGF1 are relatively well-studied, but information about the role of IGF2 in the body is still sparse. Recent discoveries called attention to emerging functions of IGF2 in the brain, where it could be involved in processes of learning and memory consolidation. It was also proposed that these functions could be mediated by the receptor for IGF2 (IGF2R). Nevertheless, little is known about the mechanism of signal transduction through this receptor. Here we produced His-tagged domain 11 (D11), an IGF2-binding element of IGF2R; we immobilized it on the solid support through a well-defined sandwich, consisting of neutravidin, biotin and synthetic anti-His-tag antibodies. Next, we prepared specifically radiolabeled [125I]-monoiodotyrosyl-Tyr2-IGF2 and optimized a sensitive and robust competitive radioligand binding assay for determination of the nanomolar binding affinities of hormones for D11 of IGF2. The assay will be helpful for the characterization of new IGF2 mutants to study the functions of IGF2R and the development of new compounds for the treatment of neurological disorders. |
| first_indexed | 2024-12-21T02:58:18Z |
| format | Article |
| id | doaj.art-c359dec76f9a4f9fbf78b085bba5e792 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T02:58:18Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-c359dec76f9a4f9fbf78b085bba5e7922022-12-21T19:18:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01159e023839310.1371/journal.pone.0238393A radioligand binding assay for the insulin-like growth factor 2 receptor.Pavlo PotalitsynIrena SelicharováKryštof SršeňJelena RadosavljevićAleš MarekKateřina NovákováJiří JiráčekLenka ŽákováInsulin-like growth factors 2 and 1 (IGF2 and IGF1) and insulin are closely related hormones that are responsible for the regulation of metabolic homeostasis, development and growth of the organism. Physiological functions of insulin and IGF1 are relatively well-studied, but information about the role of IGF2 in the body is still sparse. Recent discoveries called attention to emerging functions of IGF2 in the brain, where it could be involved in processes of learning and memory consolidation. It was also proposed that these functions could be mediated by the receptor for IGF2 (IGF2R). Nevertheless, little is known about the mechanism of signal transduction through this receptor. Here we produced His-tagged domain 11 (D11), an IGF2-binding element of IGF2R; we immobilized it on the solid support through a well-defined sandwich, consisting of neutravidin, biotin and synthetic anti-His-tag antibodies. Next, we prepared specifically radiolabeled [125I]-monoiodotyrosyl-Tyr2-IGF2 and optimized a sensitive and robust competitive radioligand binding assay for determination of the nanomolar binding affinities of hormones for D11 of IGF2. The assay will be helpful for the characterization of new IGF2 mutants to study the functions of IGF2R and the development of new compounds for the treatment of neurological disorders.https://doi.org/10.1371/journal.pone.0238393 |
| spellingShingle | Pavlo Potalitsyn Irena Selicharová Kryštof Sršeň Jelena Radosavljević Aleš Marek Kateřina Nováková Jiří Jiráček Lenka Žáková A radioligand binding assay for the insulin-like growth factor 2 receptor. PLoS ONE |
| title | A radioligand binding assay for the insulin-like growth factor 2 receptor. |
| title_full | A radioligand binding assay for the insulin-like growth factor 2 receptor. |
| title_fullStr | A radioligand binding assay for the insulin-like growth factor 2 receptor. |
| title_full_unstemmed | A radioligand binding assay for the insulin-like growth factor 2 receptor. |
| title_short | A radioligand binding assay for the insulin-like growth factor 2 receptor. |
| title_sort | radioligand binding assay for the insulin like growth factor 2 receptor |
| url | https://doi.org/10.1371/journal.pone.0238393 |
| work_keys_str_mv | AT pavlopotalitsyn aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT irenaselicharova aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT krystofsrsen aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT jelenaradosavljevic aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT alesmarek aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT katerinanovakova aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT jirijiracek aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT lenkazakova aradioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT pavlopotalitsyn radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT irenaselicharova radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT krystofsrsen radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT jelenaradosavljevic radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT alesmarek radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT katerinanovakova radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT jirijiracek radioligandbindingassayfortheinsulinlikegrowthfactor2receptor AT lenkazakova radioligandbindingassayfortheinsulinlikegrowthfactor2receptor |