Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation

Phosphorylated polypeptides represent promising biomimetic macromolecules for various regenerative applications. However to date, large-scale synthesis of phosphorylated polypeptides with controlled degrees of phosphorylation has not been achieved, restricting research in phosphorylated proteins to...

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Main Authors: Lipik, Vitali, Zhang, Lihong, Miserez, Ali
Other Authors: School of Materials Science & Engineering
Format: Article
Language:English
Published: 2014
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Online Access:https://hdl.handle.net/10356/103145
http://hdl.handle.net/10220/24413
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spelling sg-ntu-dr.10356-1031452020-06-01T10:26:37Z Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation Lipik, Vitali Zhang, Lihong Miserez, Ali School of Materials Science & Engineering School of Biological Sciences DRNTU::Science::Chemistry::Organic chemistry::Polymers Phosphorylated polypeptides represent promising biomimetic macromolecules for various regenerative applications. However to date, large-scale synthesis of phosphorylated polypeptides with controlled degrees of phosphorylation has not been achieved, restricting research in phosphorylated proteins to their central roles in biomineralization pathways. Here, we present a co-polypeptide synthesis strategy based on the Ring-Opening Polymerization (ROP) of N-carboxyanhydrides (NCAs), followed by controlled phosphorylation of serine (Ser) residues. The molecular design, including amino acid composition and molecular weight of polypeptides, mimicked the intriguing phosphorylated protein Pc-3 secreted by the sandcastle tube worm Phragmatopoma californica, which is a major constituent of the glue produced by the animal to bind hard particles together for their protective tubes. Pc-3 is comprised of mostly Ser and tyrosine (Tyr), with up to 70% of Ser residues phosphorylated into phospho-serine (pSer) giving rise to the high net negative charge of Pc-3. Three NCA monomers were synthesized, namely Ser with free –OH groups, and Ser and Tyr with protected –OH groups, and subsequently polymerized with various feeding ratios in order to obtain a broad range of final amino acid compositions. In the final step, phosphorylation targeting free –OH groups of Ser was conducted. With this strategy, the degree of phosphorylation is governed by the initial amount of unprotected –OH groups of the precursor Ser–NCA, and the final co-polypeptides contain relative amounts of Tyr and pSer that can be tailored, yielding a composition and molecular weight (MW) that closely match those of Pc-3. This control of phosphorylation leads to polypeptides exhibiting a wide range of zeta potential values between −20 and −50 mV. Using analytical assays, including Dynamic Light Scattering (DLS), Surface Plasmon Resonance (SPR), and Quartz Crystal Microbalance with Dissipation (QCM-D), we demonstrate that these phosphorylated polypeptides exhibit affinity towards divalent ions such as Ca2+, thus opening the door for their usage as scaffolds for mineralized tissue repair or as a major component of biocompatible adhesives. 2014-12-10T02:01:11Z 2019-12-06T21:06:28Z 2014-12-10T02:01:11Z 2019-12-06T21:06:28Z 2014 2014 Journal Article Lipik, V., Zhang, L., & Miserez, A. (2014). Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation. Polymer chemistry, 5(4), 1351-1361. 1759-9954 https://hdl.handle.net/10356/103145 http://hdl.handle.net/10220/24413 10.1039/c3py01118f en Polymer chemistry © 2014 The Royal Society of Chemistry.
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic DRNTU::Science::Chemistry::Organic chemistry::Polymers
spellingShingle DRNTU::Science::Chemistry::Organic chemistry::Polymers
Lipik, Vitali
Zhang, Lihong
Miserez, Ali
Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
description Phosphorylated polypeptides represent promising biomimetic macromolecules for various regenerative applications. However to date, large-scale synthesis of phosphorylated polypeptides with controlled degrees of phosphorylation has not been achieved, restricting research in phosphorylated proteins to their central roles in biomineralization pathways. Here, we present a co-polypeptide synthesis strategy based on the Ring-Opening Polymerization (ROP) of N-carboxyanhydrides (NCAs), followed by controlled phosphorylation of serine (Ser) residues. The molecular design, including amino acid composition and molecular weight of polypeptides, mimicked the intriguing phosphorylated protein Pc-3 secreted by the sandcastle tube worm Phragmatopoma californica, which is a major constituent of the glue produced by the animal to bind hard particles together for their protective tubes. Pc-3 is comprised of mostly Ser and tyrosine (Tyr), with up to 70% of Ser residues phosphorylated into phospho-serine (pSer) giving rise to the high net negative charge of Pc-3. Three NCA monomers were synthesized, namely Ser with free –OH groups, and Ser and Tyr with protected –OH groups, and subsequently polymerized with various feeding ratios in order to obtain a broad range of final amino acid compositions. In the final step, phosphorylation targeting free –OH groups of Ser was conducted. With this strategy, the degree of phosphorylation is governed by the initial amount of unprotected –OH groups of the precursor Ser–NCA, and the final co-polypeptides contain relative amounts of Tyr and pSer that can be tailored, yielding a composition and molecular weight (MW) that closely match those of Pc-3. This control of phosphorylation leads to polypeptides exhibiting a wide range of zeta potential values between −20 and −50 mV. Using analytical assays, including Dynamic Light Scattering (DLS), Surface Plasmon Resonance (SPR), and Quartz Crystal Microbalance with Dissipation (QCM-D), we demonstrate that these phosphorylated polypeptides exhibit affinity towards divalent ions such as Ca2+, thus opening the door for their usage as scaffolds for mineralized tissue repair or as a major component of biocompatible adhesives.
author2 School of Materials Science & Engineering
author_facet School of Materials Science & Engineering
Lipik, Vitali
Zhang, Lihong
Miserez, Ali
format Article
author Lipik, Vitali
Zhang, Lihong
Miserez, Ali
author_sort Lipik, Vitali
title Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
title_short Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
title_full Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
title_fullStr Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
title_full_unstemmed Synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
title_sort synthesis of biomimetic co-polypeptides with tunable degrees of phosphorylation
publishDate 2014
url https://hdl.handle.net/10356/103145
http://hdl.handle.net/10220/24413
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