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An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation

Increasing evidence suggests that signaling through the prolactin/prolactin receptor axis is important for stimulation the growth of many cancers including glioblastoma multiforme, breast and ovarian carcinoma. Efficient inhibitors of signaling have previously been developed but their applicability...

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Autores principales: Yu, Shengze, Alkharusi, Amira, Norstedt, Gunnar, Gräslund, Torbjörn
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6504076/
https://www.ncbi.nlm.nih.gov/pubmed/31063493
http://dx.doi.org/10.1371/journal.pone.0215831
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author Yu, Shengze
Alkharusi, Amira
Norstedt, Gunnar
Gräslund, Torbjörn
author_facet Yu, Shengze
Alkharusi, Amira
Norstedt, Gunnar
Gräslund, Torbjörn
author_sort Yu, Shengze
collection PubMed
description Increasing evidence suggests that signaling through the prolactin/prolactin receptor axis is important for stimulation the growth of many cancers including glioblastoma multiforme, breast and ovarian carcinoma. Efficient inhibitors of signaling have previously been developed but their applicability as cancer drugs is limited by the short in vivo half-life. In this study, we show that a fusion protein, consisting of the prolactin receptor antagonist PrlRA and an albumin binding domain for half-life extension can be expressed as inclusion bodies in Escherichia coli and efficiently refolded and purified to homogeneity. The fusion protein was found to have strong affinity for the two intended targets: the prolactin receptor (K(D) = 2.3±0.2 nM) and mouse serum albumin (K(D) = 0.38±0.01 nM). Further investigation showed that it could efficiently prevent prolactin mediated phosphorylation of STAT5 at 100 nM concentration and above, similar to the PrlRA itself, suggesting a potential as drug for cancer therapy in the future. Complexion with HSA weakened the affinity for the receptor to 21±3 nM, however the ability to prevent phosphorylation of STAT5 was still prominent. Injection into rats showed a 100-fold higher concentration in blood after 24 h compared to PrlRA itself.
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spelling pubmed-65040762019-05-09 An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation Yu, Shengze Alkharusi, Amira Norstedt, Gunnar Gräslund, Torbjörn PLoS One Research Article Increasing evidence suggests that signaling through the prolactin/prolactin receptor axis is important for stimulation the growth of many cancers including glioblastoma multiforme, breast and ovarian carcinoma. Efficient inhibitors of signaling have previously been developed but their applicability as cancer drugs is limited by the short in vivo half-life. In this study, we show that a fusion protein, consisting of the prolactin receptor antagonist PrlRA and an albumin binding domain for half-life extension can be expressed as inclusion bodies in Escherichia coli and efficiently refolded and purified to homogeneity. The fusion protein was found to have strong affinity for the two intended targets: the prolactin receptor (K(D) = 2.3±0.2 nM) and mouse serum albumin (K(D) = 0.38±0.01 nM). Further investigation showed that it could efficiently prevent prolactin mediated phosphorylation of STAT5 at 100 nM concentration and above, similar to the PrlRA itself, suggesting a potential as drug for cancer therapy in the future. Complexion with HSA weakened the affinity for the receptor to 21±3 nM, however the ability to prevent phosphorylation of STAT5 was still prominent. Injection into rats showed a 100-fold higher concentration in blood after 24 h compared to PrlRA itself. Public Library of Science 2019-05-07 /pmc/articles/PMC6504076/ /pubmed/31063493 http://dx.doi.org/10.1371/journal.pone.0215831 Text en © 2019 Yu et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yu, Shengze
Alkharusi, Amira
Norstedt, Gunnar
Gräslund, Torbjörn
An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title_full An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title_fullStr An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title_full_unstemmed An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title_short An in vivo half-life extended prolactin receptor antagonist can prevent STAT5 phosphorylation
title_sort in vivo half-life extended prolactin receptor antagonist can prevent stat5 phosphorylation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6504076/
https://www.ncbi.nlm.nih.gov/pubmed/31063493
http://dx.doi.org/10.1371/journal.pone.0215831
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