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Lysophospholipids induce fibrillation of the repeat domain of Pmel17 through intermediate core-shell structures

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Lysophospholipids induce fibrillation of the repeat domain of Pmel17 through intermediate core-shell structures. / Pedersen, Jannik Nedergaard; Jiang, Zhiping; Christiansen, Gunna; Lee, Jennifer C; Pedersen, Jan Skov; Otzen, Daniel E.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1867, No. 5, 2019, p. 519-528.

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Pedersen, Jannik Nedergaard ; Jiang, Zhiping ; Christiansen, Gunna ; Lee, Jennifer C ; Pedersen, Jan Skov ; Otzen, Daniel E. / Lysophospholipids induce fibrillation of the repeat domain of Pmel17 through intermediate core-shell structures. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2019 ; Vol. 1867, No. 5. pp. 519-528.

Bibtex

@article{fce739f4f8b649b7a791f99175c90fd5,
title = "Lysophospholipids induce fibrillation of the repeat domain of Pmel17 through intermediate core-shell structures",
abstract = "Lipids often play an important role in the initial steps of fibrillation. The melanosomal protein Pmel17 forms amyloid in vivo and contains a highly amyloidogenic Repeat domain (RPT), important for melanin biosynthesis. RPT fibrillation is influenced by two lysolipids, the anionic lysophosphatidylglycerol (LPG) and zwitterionic lysophosphatidylcholine (LPC), both present in vivo at elevated concentrations in melanosomes, organelles in which Pmel17 aggregate. Here we investigate the interaction of RPT with both LPG and LPC using small-angle X-ray scattering (SAXS), isothermal titration calorimetry (ITC), electron microscopy, fluorescence and circular dichroism (CD) spectroscopy. Under non-shaking conditions, both lipids promote fibrillation but this is driven by different interactions with RPT. Each RPT binds >40 LPG molecules but only weak interactions are seen with LPC. Above LPG's criticial micelle concentration (cmc), LPG and RPT form connected micelles where RPT binds to the surface as beads on a string with core-shell structures. Binding to LPG only induces α-helical structure well above the cmc, while LPC has no measurable effect on the protein structure. While low (but still super-cmc) concentrations of LPG strongly promote aggregation, at higher LPG concentrations (10 mM), only ~ one RPT binds per micelle, inhibiting amyloid formation. ITC and SAXS reveal some interactions between the zwitterionic lipid LPC and RPT below the cmc but little above the cmc. Nevertheless, LPC only promotes aggregation above the cmc and this process is not inhibited by high LPC concentrations, suggesting that monomers and micelles cooperate to influence amyloid formation.",
keywords = "Fibrillation, Functional amyloid, Lysolipid micelles, SAXS, ITC, pmel17",
author = "Pedersen, {Jannik Nedergaard} and Zhiping Jiang and Gunna Christiansen and Lee, {Jennifer C} and Pedersen, {Jan Skov} and Otzen, {Daniel E}",
note = "Copyright {\circledC} 2018. Published by Elsevier B.V.",
year = "2019",
doi = "10.1016/j.bbapap.2018.11.007",
language = "English",
volume = "1867",
pages = "519--528",
journal = "B B A - Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier BV",
number = "5",

}

RIS

TY - JOUR

T1 - Lysophospholipids induce fibrillation of the repeat domain of Pmel17 through intermediate core-shell structures

AU - Pedersen, Jannik Nedergaard

AU - Jiang, Zhiping

AU - Christiansen, Gunna

AU - Lee, Jennifer C

AU - Pedersen, Jan Skov

AU - Otzen, Daniel E

N1 - Copyright © 2018. Published by Elsevier B.V.

PY - 2019

Y1 - 2019

N2 - Lipids often play an important role in the initial steps of fibrillation. The melanosomal protein Pmel17 forms amyloid in vivo and contains a highly amyloidogenic Repeat domain (RPT), important for melanin biosynthesis. RPT fibrillation is influenced by two lysolipids, the anionic lysophosphatidylglycerol (LPG) and zwitterionic lysophosphatidylcholine (LPC), both present in vivo at elevated concentrations in melanosomes, organelles in which Pmel17 aggregate. Here we investigate the interaction of RPT with both LPG and LPC using small-angle X-ray scattering (SAXS), isothermal titration calorimetry (ITC), electron microscopy, fluorescence and circular dichroism (CD) spectroscopy. Under non-shaking conditions, both lipids promote fibrillation but this is driven by different interactions with RPT. Each RPT binds >40 LPG molecules but only weak interactions are seen with LPC. Above LPG's criticial micelle concentration (cmc), LPG and RPT form connected micelles where RPT binds to the surface as beads on a string with core-shell structures. Binding to LPG only induces α-helical structure well above the cmc, while LPC has no measurable effect on the protein structure. While low (but still super-cmc) concentrations of LPG strongly promote aggregation, at higher LPG concentrations (10 mM), only ~ one RPT binds per micelle, inhibiting amyloid formation. ITC and SAXS reveal some interactions between the zwitterionic lipid LPC and RPT below the cmc but little above the cmc. Nevertheless, LPC only promotes aggregation above the cmc and this process is not inhibited by high LPC concentrations, suggesting that monomers and micelles cooperate to influence amyloid formation.

AB - Lipids often play an important role in the initial steps of fibrillation. The melanosomal protein Pmel17 forms amyloid in vivo and contains a highly amyloidogenic Repeat domain (RPT), important for melanin biosynthesis. RPT fibrillation is influenced by two lysolipids, the anionic lysophosphatidylglycerol (LPG) and zwitterionic lysophosphatidylcholine (LPC), both present in vivo at elevated concentrations in melanosomes, organelles in which Pmel17 aggregate. Here we investigate the interaction of RPT with both LPG and LPC using small-angle X-ray scattering (SAXS), isothermal titration calorimetry (ITC), electron microscopy, fluorescence and circular dichroism (CD) spectroscopy. Under non-shaking conditions, both lipids promote fibrillation but this is driven by different interactions with RPT. Each RPT binds >40 LPG molecules but only weak interactions are seen with LPC. Above LPG's criticial micelle concentration (cmc), LPG and RPT form connected micelles where RPT binds to the surface as beads on a string with core-shell structures. Binding to LPG only induces α-helical structure well above the cmc, while LPC has no measurable effect on the protein structure. While low (but still super-cmc) concentrations of LPG strongly promote aggregation, at higher LPG concentrations (10 mM), only ~ one RPT binds per micelle, inhibiting amyloid formation. ITC and SAXS reveal some interactions between the zwitterionic lipid LPC and RPT below the cmc but little above the cmc. Nevertheless, LPC only promotes aggregation above the cmc and this process is not inhibited by high LPC concentrations, suggesting that monomers and micelles cooperate to influence amyloid formation.

KW - Fibrillation

KW - Functional amyloid

KW - Lysolipid micelles

KW - SAXS, ITC

KW - pmel17

UR - http://www.scopus.com/inward/record.url?scp=85057221660&partnerID=8YFLogxK

U2 - 10.1016/j.bbapap.2018.11.007

DO - 10.1016/j.bbapap.2018.11.007

M3 - Journal article

VL - 1867

SP - 519

EP - 528

JO - B B A - Proteins and Proteomics

JF - B B A - Proteins and Proteomics

SN - 1570-9639

IS - 5

ER -