Control of Protein Oligomerization and De-oligomerization on Lipid Membranes

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School of Science | Doctoral thesis (article-based) | Defence date: 2014-01-24
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Date

2013

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Degree programme

Language

en

Pages

72 + app. 105

Series

Aalto University publication series DOCTORAL DISSERTATIONS, 220/2013

Abstract

Oligomerization of protein into amyloid fibrils is central to the pathogenesis of several neurodegenerative disorders. Amyloid fibrillation and the cytotoxic actions of amyloids are membrane-associated processes. The interactions of amyloid-forming proteins with lipids at the membrane surface accelerate fibrillation and induce membrane permeabilization. Oligomerization also plays a functional role in antimicrobial defense and controls the catalytic activity of phospholipase A2 (PLA2). The protein oligomerization and amyloid formation can be modulated by heat shock protein 70 (Hsp70). Thus, the aim of the present work was to study membrane-associated protein oligomerization and its modulation by Hsp70 on the phospholipid model membrane system. Sequence analyses revealed that antimicrobial peptides (AMPs) contained sequence motifs that showed propensities for self-assembly, aggregation, and oligomerization into amyloid fibrils. The presence of such oligomerization-mediating sequences was characteristic of amyloidogenic cytotoxic proteins, including gelsolin involved in familial Finnish type amyloidosis (FAF). 1-Palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine (PoxnoPC), an oxidized phospholipid, accelerated fibrillation of the core amyloidogenic segment of gelsolin.The PoxnoPC-mediated fibrillation of gelsolin was dependent on both the concentration and the aggregation state of PoxnoPC. Fibril growth followed simple nucleation-dependent kinetics with the formation of transient prefibrillar oligomers in the lag phase. Subsequently, in order to understand the functional role of membrane-associated Hsp70, we studied lipid-Hsp70 interactions. The association of Hsp70 with phospholipid membranes was highly dependent on their lipid compositions. Hsp70 associated with phosphatidylcholine bilayers and penetrated into the hydrocarbon region. In contrast to the above data, in the presence of negatively charged phospholipids, Hsp70 bound peripherally to membrane surfacesby extended phospholipid anchorage. A specific pH-dependent association of Hsp70 with bis(monoacylglycero)phosphate, an acidic phospholipid enriched in the inner lysosomal membrane, activated lysosomal acid sphingomyelinase and promoted cell survival. We also showed that the Hsp70 sustained the hydrolytic activity of PLA2 by modulating the oligomerization and transformation of PLA2 into amyloid fibers. Hsp70 attenuated the lysophosphatidylcholine-induced inhibition and amyloid formation of PLA2 in an ATP-dependent manner. Finally, an oligomerization-mediating sequence in PLA2 was identified. Synthetic peptides corresponding to amyloidogenic, aggregation-promoting regions inhibited the hydrolytic activity of PLA2.

Description

Supervising professor

Kinnunen, Paavo K. J., Prof., Aalto University, Department of Biomedical Engineering and Computational Science, Finland

Thesis advisor

Kinnunen, Paavo K. J., Prof., Aalto University, Department of Biomedical Engineering and Computational Science, Finland

Keywords

amyloid, antimicrobial peptides, fluorescence, gelsolin, heat shock protein 70, membranes, oligomers, phospholipid, phospholipase A2, and tryptophan

Other note

Parts

  • [Publication 1]: Mahalka, A. K., Kinnunen, P. K. J., Binding of amphipathic alpha-helical antimicrobial peptides to lipid membranes, lessons from temporins B and L, BBA Biomembranes 1788 (2009) 1600-1609
  • [Publication 2]: Mahalka, A. K., Maury, C. P. J., Kinnunen, P. K. J., 1-palmitoyl-2-(9'-oxononanoyl)-sn-glycero-3-phosphocholine, an oxidized phospholipid accelerates Finnish type familial gelsolin amyloidosis in vitro, Biochemistry 50 (2011) 4877-4889
  • [Publication 3]: Mahalka, A. K., Kirkegaard, T., Jukola, L. T. I., Jäättelä, M., Kinnunen, P. K. J., Human heat shock protein 70 (Hsp70) as a peripheral membrane protein (in revision for BBA Biomembranes)
  • [Publication 4]: Kirkegaard, T., Roth, A. G., Petersen, N. H. T., Mahalka, A. K., Olsen, O. D., Moilanen, I., Zylicz, A., Knudsen, J., Sandhoff, K., Arenz, C., Kinnunen, P. K. J., Nylandsted, J., Jäättelä, M., Hsp70 stabilizes lysosomes and reverts Niemann-Pick disease-associated lysosomal pathology, Nature 463 (2010) 549-553
  • [Publication 5]: Mahalka, A. K.,*Code, C.,* Rezaijahromi, B., Kirkegaard, T., Jäättelä, M., Kinnunen, P. K. J., Activation of phospholipase A2 by Hsp70 in vitro, BBA Biomembranes 1808 (2011) 2569-2572
  • [Publication 6]: Mahalka, A. K., Kinnunen, P. K. J., Class specific peptide inhibitors for secretory phospholipases A2, Biochem Biophys Res Commun 436 (2013) 349–353
  • [Publication 7]: Parry, M. J., Alakoskela, J. M., Khandelia, H., Kumar, S.A., Jäättelä, M., Mahalka, A. K., Kinnunen, P. K. J., High-affinity small molecule- phospholipid complex formation: binding of siramesine to phosphatidic acid, J Am Chem Soc 130 (2008) 12953-12960.
  • [Publication 8]: Code, C., Mahalka, A. K., Bry, K., Kinnunen, P. K. J., Activation of phospholipase A2 by 1-palmitoyl-2-(9´-oxononanoyl)-sn-glycero-3- phosphocholine in vitro, BBA Biomembranes 1798 (2010) 1593-6000
  • [Publication 9]: Kinnunen, P. K. J., Kaarniranta, K., Mahalka, A. K., Protein-oxidized phospholipid interaction in cellular signalling: From biophysics to clinical correlations, BBA Biomembranes 1818 (2012) 2446-2455
  • [Publication 10]: Olrichs, N. K., Mahalka, A. K., Kaloyanova, D., Kinnunen, P. K. J., Helms, B., GAPR-1 forms amyloid fibrils by interaction with acidic phospholipids and inhibits Aβ aggregation. Implications for the CAP superfamily, (In revision for Amyloid)

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