Improving membrane binding as a design strategy for amphipathic peptide hormones: 2-helix variants of PYY3-36

Søren Ljungberg Pedersen; Vikram Kjøller Bhatia; Simon Jurt; Johan F. Paulsson; Maria Hauge Pedersen; Rasmus Jørgensen; Birgitte Holst; Dimitrios Stamou; Niels Vrang; Oliver Zerbe; Knud Jørgen Jensen

doi:10.1002/psc.2436

Improving membrane binding as a design strategy for amphipathic peptide hormones: 2-helix variants of PYY3-36

Søren Ljungberg Pedersen, Vikram Kjøller Bhatia, Simon Jurt, Johan F. Paulsson, Maria Hauge Pedersen, Rasmus Jørgensen, Birgitte Holst, Dimitrios Stamou, Niels Vrang, Oliver Zerbe, Knud Jørgen Jensen

7 Citations (Scopus)

Abstract

It has been hypothesized that amphipathic peptides might bind to membranes prior to activating their cognate receptors, but this has proven difficult to test. The peptide hormone PYY3-36 is believed to perform its appetite-suppressing actions through binding to hypothalamic Y2 receptors. It has been proposed that PYY3-36 via its amphipathic α-helix binds to the plasma membrane prior to receptor docking. Here, our aim was to study the implication of this hypothesis using new analogs of PYY3-36. We first studied membrane binding of PYY3-36. Next, we designed a series of PYY3-36 analogs to increase membrane-binding affinity by substituting the N-terminal segment with a de novo designed α-helical, amphipathic sequence. These 2-helix variants of PYY3-36 were assembled by solid-phase peptide synthesis. Pharmacological studies demonstrated that even though the native peptide sequence was radically changed, highly active Y2 receptor agonists were generated. A potent analog, with a Kd of 4nM for membranes, was structurally characterized by NMR in the membrane-bound state, which clearly showed that it formed the expected 2-helix. The topology of the peptide-micelle association was studied by paramagnetic relaxation enhancement using a spin label, which confirmed that the hydrophobic residues bound to the membrane. Our studies further support the hypothesis that PYY3-36 associates with the membrane and indicate that this can be used in the design of novel molecules with high receptor binding potency. These observations are likely to be generally important for peptide hormones and biopharmaceutical drugs derived from them. This new 2-helix variant of PYY3-36 will be useful as a tool compound for studying peptide-membrane interactions.

Original language	English
Journal	Journal of Peptide Science
Volume	18
Issue number	9
Pages (from-to)	579-587
Number of pages	9
ISSN	1075-2617
DOIs	https://doi.org/10.1002/psc.2436
Publication status	Published - Sept 2012

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@article{1e2737fb675a441bb683734958829b0a,

title = "Improving membrane binding as a design strategy for amphipathic peptide hormones: 2-helix variants of PYY3-36",

abstract = "It has been hypothesized that amphipathic peptides might bind to membranes prior to activating their cognate receptors, but this has proven difficult to test. The peptide hormone PYY3-36 is believed to perform its appetite-suppressing actions through binding to hypothalamic Y2 receptors. It has been proposed that PYY3-36 via its amphipathic α-helix binds to the plasma membrane prior to receptor docking. Here, our aim was to study the implication of this hypothesis using new analogs of PYY3-36. We first studied membrane binding of PYY3-36. Next, we designed a series of PYY3-36 analogs to increase membrane-binding affinity by substituting the N-terminal segment with a de novo designed α-helical, amphipathic sequence. These 2-helix variants of PYY3-36 were assembled by solid-phase peptide synthesis. Pharmacological studies demonstrated that even though the native peptide sequence was radically changed, highly active Y2 receptor agonists were generated. A potent analog, with a Kd of 4nM for membranes, was structurally characterized by NMR in the membrane-bound state, which clearly showed that it formed the expected 2-helix. The topology of the peptide-micelle association was studied by paramagnetic relaxation enhancement using a spin label, which confirmed that the hydrophobic residues bound to the membrane. Our studies further support the hypothesis that PYY3-36 associates with the membrane and indicate that this can be used in the design of novel molecules with high receptor binding potency. These observations are likely to be generally important for peptide hormones and biopharmaceutical drugs derived from them. This new 2-helix variant of PYY3-36 will be useful as a tool compound for studying peptide-membrane interactions.",

author = "Pedersen, {S{\o}ren Ljungberg} and Bhatia, {Vikram Kj{\o}ller} and Simon Jurt and Paulsson, {Johan F.} and Pedersen, {Maria Hauge} and Rasmus J{\o}rgensen and Birgitte Holst and Dimitrios Stamou and Niels Vrang and Oliver Zerbe and Jensen, {Knud J{\o}rgen}",

year = "2012",

month = sep,

doi = "10.1002/psc.2436",

language = "English",

volume = "18",

pages = "579--587",

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T1 - Improving membrane binding as a design strategy for amphipathic peptide hormones

T2 - 2-helix variants of PYY3-36

AU - Pedersen, Søren Ljungberg

AU - Bhatia, Vikram Kjøller

AU - Jurt, Simon

AU - Paulsson, Johan F.

AU - Pedersen, Maria Hauge

AU - Jørgensen, Rasmus

AU - Holst, Birgitte

AU - Stamou, Dimitrios

AU - Vrang, Niels

AU - Zerbe, Oliver

AU - Jensen, Knud Jørgen

PY - 2012/9

Y1 - 2012/9

N2 - It has been hypothesized that amphipathic peptides might bind to membranes prior to activating their cognate receptors, but this has proven difficult to test. The peptide hormone PYY3-36 is believed to perform its appetite-suppressing actions through binding to hypothalamic Y2 receptors. It has been proposed that PYY3-36 via its amphipathic α-helix binds to the plasma membrane prior to receptor docking. Here, our aim was to study the implication of this hypothesis using new analogs of PYY3-36. We first studied membrane binding of PYY3-36. Next, we designed a series of PYY3-36 analogs to increase membrane-binding affinity by substituting the N-terminal segment with a de novo designed α-helical, amphipathic sequence. These 2-helix variants of PYY3-36 were assembled by solid-phase peptide synthesis. Pharmacological studies demonstrated that even though the native peptide sequence was radically changed, highly active Y2 receptor agonists were generated. A potent analog, with a Kd of 4nM for membranes, was structurally characterized by NMR in the membrane-bound state, which clearly showed that it formed the expected 2-helix. The topology of the peptide-micelle association was studied by paramagnetic relaxation enhancement using a spin label, which confirmed that the hydrophobic residues bound to the membrane. Our studies further support the hypothesis that PYY3-36 associates with the membrane and indicate that this can be used in the design of novel molecules with high receptor binding potency. These observations are likely to be generally important for peptide hormones and biopharmaceutical drugs derived from them. This new 2-helix variant of PYY3-36 will be useful as a tool compound for studying peptide-membrane interactions.

AB - It has been hypothesized that amphipathic peptides might bind to membranes prior to activating their cognate receptors, but this has proven difficult to test. The peptide hormone PYY3-36 is believed to perform its appetite-suppressing actions through binding to hypothalamic Y2 receptors. It has been proposed that PYY3-36 via its amphipathic α-helix binds to the plasma membrane prior to receptor docking. Here, our aim was to study the implication of this hypothesis using new analogs of PYY3-36. We first studied membrane binding of PYY3-36. Next, we designed a series of PYY3-36 analogs to increase membrane-binding affinity by substituting the N-terminal segment with a de novo designed α-helical, amphipathic sequence. These 2-helix variants of PYY3-36 were assembled by solid-phase peptide synthesis. Pharmacological studies demonstrated that even though the native peptide sequence was radically changed, highly active Y2 receptor agonists were generated. A potent analog, with a Kd of 4nM for membranes, was structurally characterized by NMR in the membrane-bound state, which clearly showed that it formed the expected 2-helix. The topology of the peptide-micelle association was studied by paramagnetic relaxation enhancement using a spin label, which confirmed that the hydrophobic residues bound to the membrane. Our studies further support the hypothesis that PYY3-36 associates with the membrane and indicate that this can be used in the design of novel molecules with high receptor binding potency. These observations are likely to be generally important for peptide hormones and biopharmaceutical drugs derived from them. This new 2-helix variant of PYY3-36 will be useful as a tool compound for studying peptide-membrane interactions.

U2 - 10.1002/psc.2436

DO - 10.1002/psc.2436

M3 - Journal article

C2 - 22865741

SN - 1075-2617

VL - 18

SP - 579

EP - 587

JO - Journal of Peptide Science

JF - Journal of Peptide Science

IS - 9

ER -

Improving membrane binding as a design strategy for amphipathic peptide hormones: 2-helix variants of PYY3-36

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