Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1

Michael Szardenings (Corresponding Author), Susanna Törnroth, Felikss Mutulis, Ruta Muceniece, Kari Keinänen, Arja Kuusinen, Jarl Wikberg

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

A phage display system for the selection of peptides binding to heterologously expressed human melanocortin receptor 1 on the surface of insect cells has been established. It could be shown that phage particles displaying the natural ligand α-melanocyte-stimulating hormone bind selectively to cells expressing this receptor and that these phages exhibit biological activity on mouse B16F1 melanoma cells. Insect cells were superior to other cell lines tested and have been used to select binders from a small library, in which critical determinants (Phe7-Arg8-Trp9) were kept, whereas the flanking regions where allowed to variate freely. One peptide displaying little similarity with native hormone was found that binds to the receptor also in its free form with an affinity of 7 nm. It showed a remarkable selectivity for this receptor, because it binds to the other melanocortin receptor subtypes with a maximum affinity of 21 μm. This is the first time phage display has been used successfully with G-protein-coupled receptors lacking an extracellular binding domain.

Original languageEnglish
Pages (from-to)27943 - 27948
Number of pages6
JournalJournal of Biological Chemistry
Volume272
Issue number44
DOIs
Publication statusPublished - 1997
MoE publication typeA1 Journal article-refereed

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Receptor, Melanocortin, Type 1
Bacteriophages
Display devices
Peptides
Melanocortin Receptors
Cells
Melanocyte-Stimulating Hormones
Virus Receptors
Insects
G-Protein-Coupled Receptors
Bioactivity
Binders
Hormones
Libraries
Ligands
Melanoma
Cell Line

Cite this

Szardenings, M., Törnroth, S., Mutulis, F., Muceniece, R., Keinänen, K., Kuusinen, A., & Wikberg, J. (1997). Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1. Journal of Biological Chemistry, 272(44), 27943 - 27948. https://doi.org/10.1074/jbc.272.44.27943
Szardenings, Michael ; Törnroth, Susanna ; Mutulis, Felikss ; Muceniece, Ruta ; Keinänen, Kari ; Kuusinen, Arja ; Wikberg, Jarl. / Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 44. pp. 27943 - 27948.
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abstract = "A phage display system for the selection of peptides binding to heterologously expressed human melanocortin receptor 1 on the surface of insect cells has been established. It could be shown that phage particles displaying the natural ligand α-melanocyte-stimulating hormone bind selectively to cells expressing this receptor and that these phages exhibit biological activity on mouse B16F1 melanoma cells. Insect cells were superior to other cell lines tested and have been used to select binders from a small library, in which critical determinants (Phe7-Arg8-Trp9) were kept, whereas the flanking regions where allowed to variate freely. One peptide displaying little similarity with native hormone was found that binds to the receptor also in its free form with an affinity of 7 nm. It showed a remarkable selectivity for this receptor, because it binds to the other melanocortin receptor subtypes with a maximum affinity of 21 μm. This is the first time phage display has been used successfully with G-protein-coupled receptors lacking an extracellular binding domain.",
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Szardenings, M, Törnroth, S, Mutulis, F, Muceniece, R, Keinänen, K, Kuusinen, A & Wikberg, J 1997, 'Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1', Journal of Biological Chemistry, vol. 272, no. 44, pp. 27943 - 27948. https://doi.org/10.1074/jbc.272.44.27943

Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1. / Szardenings, Michael (Corresponding Author); Törnroth, Susanna; Mutulis, Felikss; Muceniece, Ruta; Keinänen, Kari; Kuusinen, Arja; Wikberg, Jarl.

In: Journal of Biological Chemistry, Vol. 272, No. 44, 1997, p. 27943 - 27948.

Research output: Contribution to journalArticleScientificpeer-review

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AB - A phage display system for the selection of peptides binding to heterologously expressed human melanocortin receptor 1 on the surface of insect cells has been established. It could be shown that phage particles displaying the natural ligand α-melanocyte-stimulating hormone bind selectively to cells expressing this receptor and that these phages exhibit biological activity on mouse B16F1 melanoma cells. Insect cells were superior to other cell lines tested and have been used to select binders from a small library, in which critical determinants (Phe7-Arg8-Trp9) were kept, whereas the flanking regions where allowed to variate freely. One peptide displaying little similarity with native hormone was found that binds to the receptor also in its free form with an affinity of 7 nm. It showed a remarkable selectivity for this receptor, because it binds to the other melanocortin receptor subtypes with a maximum affinity of 21 μm. This is the first time phage display has been used successfully with G-protein-coupled receptors lacking an extracellular binding domain.

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Szardenings M, Törnroth S, Mutulis F, Muceniece R, Keinänen K, Kuusinen A et al. Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1. Journal of Biological Chemistry. 1997;272(44):27943 - 27948. https://doi.org/10.1074/jbc.272.44.27943