Prof. dr. Antoinette Killian

Model membranes and protein-lipid interactions

Membranes are vital for almost all cellular functions. Their structure, function and organization are largely determined by the molecular interactions between membrane constituents, i.e. proteins and lipids within and at membranes. These interactions can be conveniently studied by making use of model membrane systems.
Main current research lines involving model membrane systems are the following:

1. Nanodiscs

We explore the use of amphipathic copolymers of styrene and maleic acid (SMA) as novel tools in membrane research. SMA copolymers are able to solubilize membranes in the form of polymer-bounded nanodiscs, allowing purification and characterization of membrane proteins directly in their native environment without the use of detergent. We use the SMA technology to solubilize, purify, and characterize membrane proteins in their native lipid environment, and we investigate the mode of action of the polymer using model membrane systems. In addition, we synthesize related amphipathic copolymers and characterize their activity in model membrane systems and in native membranes. Finally, we explore new applications of SMA in membrane research, in particular with respect to studying protein/lipid interactions.

Schematic representation of ways of reconstituting membrane proteins in a bilayer environment

Schematic representation of ways of reconstituting membrane proteins in a bilayer environment after purification in micelles (black arrows) versus direct solubilization of membrane proteins from their native environment in the form of native nanodiscs (red arrow). Adapted from: Dörr JM et al. (2016) Eur Biophys J. 45, 3-21.
DOI:10.1007/s00249-015-1093-y

2. Amyloid membrane interactions

This research line concerns the interaction of membranes with amyloid forming proteins, which have been associated with a wide range of diseases. Membranes play an important role in the lethal action of these proteins. On one hand they can catalyze fibril formation of amyloid proteins that bind to the membrane surface. On the other hand, fibril formation on the membrane surface can cause membrane damage and the fibrils can take up lipids from the membrane. In vivo these processes may lead to cell death.
We use commercially available synthetic peptides to study the interactions of amyloid forming proteins with (model)membranes and to test effects of inhibitors. For this we mostly use biophysical and biochemical techniques.
The amyloid protein that we focus on is the islet amyloid polypeptide (IAPP) involved in type II diabetes. This is particularly relevant in view of the worldwide increasing prevalence of obesity, which is an important risk factor in development of diabetes.

Models for the hIAPP-membrane interaction in relation to membrane damage and hIAPP cytotoxicity

Models for the hIAPP-membrane interaction in relation to membrane damage and hIAPP cytotoxicity. Adapted from: Khemtémourian L et al. (2008) Exp Diabetes Res. 421287.
DOI: 10.1155/2008/421287

Recent Publications

Luteijn RD, Praest P, Frank Thiele F, Sadasivam SM, Singethan K, Drijfhout JW, Bach C, de Boer SM, Lebbink RJ, Tao S, Helfer M, Bach NC, Protzer U, Costa AI, Killian JA, Drexler I, Wiertz EJHJ.
A broad-spectrum antiviral peptide blocks infection of viruses by binding to phosphatidylserine in the viral envelope. Cells 2020; 9,1989. PMID: 32872420. DOI:10.3390/cells9091989.

Cunningham RD, Kopf AH, Elenbaas BOW, Staal BBP, Pfukwa R, Killian JA, Klumperman B.
Iterative RAFT-Mediated Copolymerization of Styrene and Maleic Anhydride toward Sequence- and Length-Controlled Copolymers and Their Applications for Solubilizing Lipid Membranes. Biomacromolecules. 2020; 10;21(8):3287-3300. PMID: 32672942. DOI:10.1021/acs.biomac.0c00736.

Kopf AH, Dörr JM, Koorengevel MC, Antoniciello F, Jahn H, Killian JA.
Factors influencing the solubilization of membrane proteins from Escherichia coli membranes by styrene-maleic acid copolymers. Biochim Biophys Acta Biomembr. 2020; 1862(2):183125. DOI:10.1016/j.bbamem.2019.183125.

Saravanan MS, Ryazanov S, Leonov A, Nicolai J, Praest P, Giese A, Winter R, Khemtemourian L, Griesinger C, Killian JA.
The small molecule inhibitor anle145c thermodynamically traps human islet amyloid peptide in the form of non-cytotoxic oligomers. Sci Rep. 2019 Dec 13;9(1):19023. doi: 10.1038/s41598-019-54919-z.

Domínguez Pardo JJ, van Walree CA, Egmond MR, Koorengevel MC, Killian JA.
Nanodiscs bounded by styrene-maleic acid allow trans-cis isomerization of enclosed photoswitches of azobenzene labeled lipids Chem Phys Lipids. 2019 Feb 16. pii: S0009-3084(18)30245-7. doi: 10.1016/j.chemphyslip.2019.02.002. [Epub ahead of print

Kopf AH, Koorengevel MC, van Walree CA, Dafforn TR, Killian JA.
A simple and convenient method for the hydrolysis of styrene-maleic anhydride copolymers to styrene-maleic acid copolymers. Chem Phys Lipids. 2019 Jan;218:85-90. doi: 10.1016/j.chemphyslip.2018.11.011. Epub 2018 Dec 1

Liu Y, Moura ECCM, Dörr JM, Scheidelaar S, Heger M, Egmond MR, Killian JA, Mohammadi T, Breukink E.
Bacillus subtilis MraY in detergent-free system of nanodiscs wrapped by styrene-maleic acid copolymers.PLoS One. 2018 Nov 5;13(11):e0206692. doi: 10.1371/journal.pone.0206692. eCollection 2018

Dominguez Pardo JJ, Koorengevel MC, Uwugiaren N, Weijers J, Kopf AH, Jahn H, van Walree CA, van Steenbergen MJ, Killian JA
Membrane Solubilization by Styrene-Maleic Acid Copolymers: Delineating the Role of Polymer Length. Biophys J. 2018 Jul3; 115(1):129-138 doi 10.1016/bpj 2018.05.032

Hoffmann ARF, Saravanan MS, Lequin O, Killian JA, Khemtemourian L
A single mutation on the human amyloid polypeptide modulates fibril growth and affects the mechanism of amyloid-imduced membrane damage. Biochim Biophys Acta 2018 Feb 27 doi 10.1016/j.bbamem.2018.02.018

Dominguez Pardo JJ, Dörr JM, Renne MF, Ould-Braham T, Koorengevel MC, van Steenbergen MJ, Killian JA
Thermotropic properties of phosphatidylcholine nanodiscs bounded by styrene-maleic acid copolymers. J. Chem Phys Lip 2017 Nov;208:58-64. doi: 10.1016/j.chemphyslop.2017.08.0101

Dörr JM, van Coevorden MH, Hoogenraad CC, Killian JA
Solubilization of human cells by the styrene-maleic acid copolymer: insights from fluorescence microscopy. Biochim Biophys Acta 2017 Nov;1859(11):2155-2160. doi: 10.1016/j.bbamem.2017.08.010.

Khemtemourian L, Guillemain G, Foufelle F, Killian JA
Residue specific effects of human islet polypeptide amyloid on self-assembly and on cell toxicity. Biochimie. 2017 Aug; pii: S0300-9084(17)30186-4. doi: 10.1016/j.biochi.2017.07.015. PMID: 28778718

Swainsbury DJK, Scheidelaar S, Foster N, van Grondelle R, Killian JA, Jones MR
The effectiveness of styrene-maleic acid (SMA) copolymers for solubilisation of integral membrane proteins from SMA-accessible and SMA-resistant membranes. Biochim Biophys Acta. 2017 Jul 24. pii: S0005-2736(17)30236-5. doi: 10.1016/j.bbamem.2017.07.011. PMID: 28751090. Epub 2017 Jul 24

Bersch B, Dörr JM, Hessel A, Killian JA, Schanda P
Proton-Detected Solid-State NMR Spectroscopy of a Zinc Diffusion Facilitator Protein in Native Nanodiscs. Angew Chem Int Ed Engl. 2017 Jan 27. doi: 10.1002/anie.201610441. PMID: 28128538

Dominguez Pardo JJ, Dörr JM, Iyer A, Cox RC, Scheidelaar S, Koorengevel MC, Subramaniam V, Killian JA
Solubilization of lipids and lipid phases by the styrene-maleic acid copolymer. Eur Biophys J. 2017 Jan;46(1):91-101. doi: 10.1007/s00249-016-1181-7. PMID: 27815573

Scheidelaar S, Koorengevel MC, van Waalree CA, Dominguez JJ, Dörr JM, Killian JA
Effect of Polymer Composition and pH on Membrane Solubilization by Styrene-Maleic Acid Copolymers. Biphysical Journal 2016 Nov 1; 111(9): 1974-1986

Gruber JM, Scheidelaar S, van Roon H, Dekker JP, Killian JA, van Grondelle R
Photophysics in single light-harvesting complexes II: from micelle to native nanodisks Proc. of SPIE 2016 Vol. 9714 97140A-1

Dörr JM, Scheidelaar S, Koorengevel MC, Dominguez JJ, Schäfer M, van Walree CA, Killian JA
The styrene-maleic acid copolymer: a versatile tool in membrane research Eur Biophys J. 2016 Jan;45(1):3-21.

Furse S, Wienk H, Boelens R, de Kroon AI, Killian JA
E. coli MG1655 modulates its phospholipid composition through the cell cycle FEBS let 2015 Sep 14;589(19 PT B):2726-30

Cybulski LE, Ballering J, Moussatova A, Inda ME, Vazquez DB, Wassenaar TA, de Mendoza D, Tieleman DP, Killian JA
Activation of the bacterial thermosensor DesK involves a serine zipper dimerization motif that is modulated by bilayer thickness.Proc Natl Acad Sci U S A. 2015 ; 112(20):6353-8

Furse S, Egmond MR, Killian JA
Isolation of lipids from biological samples. Mol Membr Biol. 2015 32(3): 55-64

Scheidelaar S, Koorengevel MC, Dominguez Pardo JD, Meeldijk JD, Breukink E, Killian JA
Molecular model for the solubilization of membranes into nanodisks by styrene maleic Acid copolymers. Biophys J. 2015 Jan 20;108(2):279-90

Dörr JM, Koorengevel MC, Schäfer M, Prokofyev AV, Scheidelaar S, van der Cruijsen EA, Dafforn TR, Baldus M, Killian JA.
Detergent-free isolation, characterization, and functional reconstitution of a tetrameric K+ channel: the power of native nanodiscs. Proc Natl Acad Sci USA. 2014 Dec 30;111(52):18607-12. doi: 10.1073/pnas.1416205112. PMID: 25512535

Swainsbury DJ, Scheidelaar S, van Grondelle R, Killian JA, Jones MR.
Bacterial reaction centers purified with styrene maleic acid copolymer retain native membrane functional properties and display enhanced stability. Angew Chem Int Ed Engl. 2014 Oct 27;53(44):11803-7. doi: 10.1002/anie.201406412. Epub 2014 Sep 11. PMID: 25212490