Progressive mitochondrial dysfunction due to the accumulation of amyloid beta (Aβ) peptide within the mitochondrial matrix represents one of the key characteristics of Alzheimer's disease (AD) and appears already in its early stages. Inside the mitochondria, Aβ interacts with a number of biomolecules, including cyclophilin D (cypD) and 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), and affects their physiological functions. However, despite intensive ongoing research, the exact mechanisms through which Aβ impairs mitochondrial functions remain to be explained. In this work, we studied the interactions of Aβ with cypD and 17β-HSD10 in vitro using the surface plasmon resonance (SPR) method and determined the kinetic parameters (association and dissociation rates) of these interactions. This is the first work which determines all these parameters under the same conditions, thus, enabling direct comparison of relative affinities of Aβ to its mitochondrial binding partners. Moreover, we used the determined characteristics of the individual interactions to simulate the concurrent interactions of Aβ with cypD and 17β-HSD10 in different model situations associated with the progression of AD. This study not only advances the understanding of Aβ-induced processes in mitochondria during AD, but it also provides a new perspective on research into complex multi-interaction biomolecular processes in general.

Institute of Photonics and Electronics of the Czech Academy of Sciences Chaberská 1014 57 182 51 Prague Czech Republic

National Institute of Mental Health Topolová 748 250 67 Klecany Czech Republic

Zobrazit více v PubMed

Gulisano W., Maugeri D., Baltrons M.A., Fa M., Amato A., Palmeri A., D’Adamio L., Grassi C., Devanand D.P., Honig L.S., et al. Role of Amyloid-beta and Tau Proteins in Alzheimer’s Disease: Confuting the Amyloid Cascade. J. Alzheimer’s Dis. JAD. 2018;64:S611–S631. doi: 10.3233/JAD-179935. PubMed DOI PMC

Reddy P.H., Tripathi R., Troung Q., Tirumala K., Reddy T.P., Anekonda V., Shirendeb U.P., Calkins M.J., Reddy A.P., Mao P., et al. Abnormal mitochondrial dynamics and synaptic degeneration as early events in Alzheimer’s disease: Implications to mitochondria-targeted antioxidant therapeutics. Biochim. Biophys. Acta (BBA) Mol. Basis Dis. 2012;1822:639–649. doi: 10.1016/j.bbadis.2011.10.011. PubMed DOI PMC

Du H., Guo L., Yan S.S. Synaptic Mitochondrial Pathology in Alzheimer’s Disease. Antioxid. Redox Signal. 2012;16:1467–1475. doi: 10.1089/ars.2011.4277. PubMed DOI PMC

Swerdlow R.H., Burns J.M., Khan S.M. The Alzheimer’s disease mitochondrial cascade hypothesis: Progress and perspectives. Biochim. Biophys. Acta (BBA) Mol. Basis Dis. 2014;1842:1219–1231. doi: 10.1016/j.bbadis.2013.09.010. PubMed DOI PMC

Manczak M., Anekonda T.S., Henson E., Park B.S., Quinn J., Reddy P.H. Mitochondria are a direct site of A beta accumulation in Alzheimer’s disease neurons: Implications for free radical generation and oxidative damage in disease progression. Hum. Mol. Genet. 2006;15:1437–1449. doi: 10.1093/hmg/ddl066. PubMed DOI

Suzuki N., Cheung T., Cai X., Odaka A., Otvos L., Eckman C., Golde T., Younkin S. An increased percentage of long amyloid beta protein secreted by familial amyloid beta protein precursor (beta APP717) mutants. Science. 1994;264:1336–1340. doi: 10.1126/science.8191290. PubMed DOI

Pearson H.A., Peers C. Physiological roles for amyloid β peptides. J. Physiol. 2006;575:5–10. doi: 10.1113/jphysiol.2006.111203. PubMed DOI PMC

Murphy M.P., LeVine H., 3rd Alzheimer’s disease and the amyloid-beta peptide. J. Alzheimer’s Dis. JAD. 2010;19:311–323. doi: 10.3233/JAD-2010-1221. PubMed DOI PMC

Wang Y.-J., Zhou H.-D., Zhou X.-F. Clearance of amyloid-beta in Alzheimer’s disease: Progress, problems and perspectives. Drug Discov. Today. 2006;11:931–938. doi: 10.1016/j.drudis.2006.08.004. PubMed DOI

Reddy P.H. Amyloid beta, mitochondrial structural and functional dynamics in Alzheimer’s disease. Exp. Neurol. 2009;218:286–292. doi: 10.1016/j.expneurol.2009.03.042. PubMed DOI PMC

Crouch P.J., Harding S.-M.E., White A.R., Camakaris J., Bush A.I., Masters C.L. Mechanisms of Aβ mediated neurodegeneration in Alzheimer’s disease. Int. J. Biochem. Cell Biol. 2008;40:181–198. doi: 10.1016/j.biocel.2007.07.013. PubMed DOI

Lührs T., Ritter C., Adrian M., Riek-Loher D., Bohrmann B., Döbeli H., Schubert D., Riek R. 3D structure of Alzheimer’s amyloid-β(1–42) fibrils. Proc. Natl. Acad. Sci. USA. 2005;102:17342–17347. doi: 10.1073/pnas.0506723102. PubMed DOI PMC

Garai K., Frieden C. Quantitative analysis of the time course of Aβ oligomerization and subsequent growth steps using tetramethylrhodamine-labeled Aβ. Proc. Natl. Acad. Sci. USA. 2013;110:3321–3326. doi: 10.1073/pnas.1222478110. PubMed DOI PMC

Yan Y., Wang C. Aβ42 is More Rigid than Aβ40 at the C Terminus: Implications for Aβ Aggregation and Toxicity. J. Mol. Biol. 2006;364:853–862. doi: 10.1016/j.jmb.2006.09.046. PubMed DOI

Hansson Petersen C.A., Alikhani N., Behbahani H., Wiehager B., Pavlov P.F., Alafuzoff I., Leinonen V., Ito A., Winblad B., Glaser E., et al. The amyloid beta-peptide is imported into mitochondria via the TOM import machinery and localized to mitochondrial cristae. Proc. Natl. Acad. Sci. USA. 2008;105:13145–13150. doi: 10.1073/pnas.0806192105. PubMed DOI PMC

Benek O., Aitken L., Hroch L., Kuca K., Gunn-Moore F., Musilek K. A Direct Interaction between Mitochondrial Proteins and Amyloid-beta Peptide and its Significance for the Progression and Treatment of Alzheimer’s Disease. Curr. Med. Chem. 2015;22:1056–1085. doi: 10.2174/0929867322666150114163051. PubMed DOI

Muirhead K.E., Borger E., Aitken L., Conway S.J., Gunn-Moore F.J. The consequences of mitochondrial amyloid beta-peptide in Alzheimer’s disease. Biochem. J. 2010;426:255–270. doi: 10.1042/BJ20091941. PubMed DOI

Pagani L., Eckert A. Amyloid-Beta interaction with mitochondria. Int. J. Alzheimer’s Dis. 2011;2011 doi: 10.4061/2011/925050. PubMed DOI PMC

Readnower R.D., Sauerbeck A.D., Sullivan P.G. Mitochondria, Amyloid β, and Alzheimer’s Disease. Int. J. Alzheimer’s Dis. 2011;2011 doi: 10.4061/2011/104545. PubMed DOI PMC

Yan S.D., Stern D.M. Mitochondrial dysfunction and Alzheimer’s disease: Role of amyloid-β peptide alcohol dehydrogenase (ABAD) Int. J. Exp. Pathol. 2005;86:161–171. doi: 10.1111/j.0959-9673.2005.00427.x. PubMed DOI PMC

Singh P., Suman S., Chandna S., Das T.K. Possible role of amyloid-beta, adenine nucleotide translocase and cyclophilin-D interaction in mitochondrial dysfunction of Alzheimer’s disease. Bioinformation. 2009;3:440–445. doi: 10.6026/97320630003440. PubMed DOI PMC

Berridge M.J. Calcium hypothesis of Alzheimer’s disease. Pflügers Arch. Eur. J. Physiol. 2010;459:441–449. doi: 10.1007/s00424-009-0736-1. PubMed DOI

Du H., Yan S.S. Mitochondrial permeability transition pore in Alzheimer’s disease: Cyclophilin D and amyloid beta. Biochim. Biophys. Acta (BBA) Mol. Basis Dis. 2010;1802:198–204. doi: 10.1016/j.bbadis.2009.07.005. PubMed DOI PMC

Du Yan S., Fu J., Soto C., Chen X., Zhu H., Al-Mohanna F., Collison K., Zhu A., Stern E., Saido T., et al. An intracellular protein that binds amyloid-β peptide and mediates neurotoxicity in Alzheimer’s disease. Nature. 1997;389:689–695. doi: 10.1038/39522. PubMed DOI

Du Yan S., Shi Y., Zhu A., Fu J., Zhu H., Zhu Y., Gibson L., Stern E., Collison K., Al-Mohanna F., et al. Role of ERAB/l-3-Hydroxyacyl-coenzyme A Dehydrogenase Type II Activity in Aβ-induced Cytotoxicity. J. Biol. Chem. 1999;274:2145–2156. doi: 10.1074/jbc.274.4.2145. PubMed DOI

Lustbader J.W., Cirilli M., Lin C., Xu H.W., Takuma K., Wang N., Caspersen C., Chen X., Pollak S., Chaney M., et al. ABAD Directly Links Aß to Mitochondrial Toxicity in Alzheimer’s Disease. Science. 2004;304:448–452. doi: 10.1126/science.1091230. PubMed DOI

Du H., Guo L., Fang F., Chen D., Sosunov A.A., McKhann G.M., Yan Y., Wang C., Zhang H., Molkentin J.D., et al. Cyclophilin D deficiency attenuates mitochondrial and neuronal perturbation and ameliorates learning and memory in Alzheimer’s disease. Nat. Med. 2008;14:1097–1105. doi: 10.1038/nm.1868. PubMed DOI PMC

Hemmerová E., Špringer T., Krištofiková Z., Homola J. In vitro study of interaction of 17β-hydroxysteroid dehydrogenase type 10 and cyclophilin D and its potential implications for Alzheimer’s disease. Sci. Rep. 2019;9:1–12. doi: 10.1038/s41598-019-53157-7. PubMed DOI PMC

Yan Y., Liu Y., Sorci M., Belfort G., Lustbader J.W., Yan S.S., Wang C. Surface Plasmon Resonance and Nuclear Magnetic Resonance Studies of ABAD−Aβ Interaction. Biochemistry. 2007;46:1724–1731. doi: 10.1021/bi061314n. PubMed DOI

Aitken L., Quinn S.D., Perez-Gonzalez D.C., Samuel I.D.W., Penedo-Esteiro J.C., Gunn-Moore F.J. Morphology-specific inhibition of β-amyloid aggregates by 17β-hydroxysteroid dehydrogenase type 10. ChemBioChem. 2016;17:1029–1037. doi: 10.1002/cbic.201600081. PubMed DOI

Špringer T., Piliarik M., Homola J. Surface plasmon resonance sensor with dispersionless microfluidics for direct detection of nucleic acids at the low femtomole level. Sens. Actuators B Chem. 2010;145:588–591. doi: 10.1016/j.snb.2009.11.018. DOI

Špringer T., Chadtová Song X., Ermini M.L., Lamačová J., Homola J. Functional gold nanoparticles for optical affinity biosensing. Anal. Bioanal. Chem. 2017;409:4087–4097. doi: 10.1007/s00216-017-0355-1. PubMed DOI

Hou L., Shao H., Zhang Y., Li H., Menon N.K., Neuhaus E.B., Brewer J.M., Byeon I.-J.L., Ray D.G., Vitek M.P., et al. Solution NMR Studies of the Aβ(1−40) and Aβ(1−42) Peptides Establish that the Met35 Oxidation State Affects the Mechanism of Amyloid Formation. J. Am. Chem. Soc. 2004;126:1992–2005. doi: 10.1021/ja036813f. PubMed DOI

Broersen K., Jonckheere W., Rozenski J., Vandersteen A., Pauwels K., Pastore A., Rousseau F., Schymkowitz J. A standardized and biocompatible preparation of aggregate-free amyloid beta peptide for biophysical and biological studies of Alzheimer’s disease. Protein Eng. Des. Sel. 2011;24:743–750. doi: 10.1093/protein/gzr020. PubMed DOI

Bartolini M., Naldi M., Fiori J., Valle F., Biscarini F., Nicolau D.V., Andrisano V. Kinetic characterization of amyloid-beta 1–42 aggregation with a multimethodological approach. Anal. Biochem. 2011;414:215–225. doi: 10.1016/j.ab.2011.03.020. PubMed DOI

Bruggink K.A., Muller M., Kuiperij H.B., Verbeek M.M. Methods for analysis of amyloid-beta aggregates. J. Alzheimer’s Dis. JAD. 2012;28:735–758. doi: 10.3233/JAD-2011-111421. PubMed DOI

Aguilar M.-I., Small D.H. Surface plasmon resonance for the analysis of β-amyloid interactions and fibril formation in alzheimer’s disease research. Neurotox. Res. 2005;7:17–27. doi: 10.1007/BF03033773. PubMed DOI

Kaasik A., Safiulina D., Zharkovsky A., Veksler V. Regulation of mitochondrial matrix volume. Am. J. Physiol. Cell Physiol. 2007;292:C157–C163. doi: 10.1152/ajpcell.00272.2006. PubMed DOI

Laskowski M., Augustynek B., Kulawiak B., Koprowski P., Bednarczyk P., Jarmuszkiewicz W., Szewczyk A. What do we not know about mitochondrial potassium channels? Biochim. Biophys. Acta (BBA) Bioenerg. 2016;1857:1247–1257. doi: 10.1016/j.bbabio.2016.03.007. PubMed DOI

Bradshaw P.C., Pfeiffer D.R. Release of Ca2+ and Mg2+ from yeast mitochondria is stimulated by increased ionic strength. BMC Biochem. 2006;7:1–12. doi: 10.1186/1471-2091-7-4. PubMed DOI PMC

Gout E., Rebeille F., Douce R., Bligny R. Interplay of Mg2+, ADP, and ATP in the cytosol and mitochondria: Unravelling the role of Mg2+ in cell respiration. Proc. Natl. Acad. Sci. USA. 2014;111:E4560–E4567. doi: 10.1073/pnas.1406251111. PubMed DOI PMC

van der Anton Merwe P., Neil Barclay A. Transient intercellular adhesion: The importance of weak protein-protein interactions. Trends Biochem. Sci. 1994;19:354–358. doi: 10.1016/0968-0004(94)90109-0. PubMed DOI

Krištofiková Z., Špringer T., Gedeonová E., Hofmannová A., Říčný J., Hromadková L., Vyhnálek M., Laczo J., Nikolai T., Hort J., et al. Interactions of 17β-Hydroxysteroid Dehydrogenase Type 10 and Cyclophilin D in Alzheimer’s Disease. Neurochem. Res. 2020 doi: 10.1007/s11064-020-02970-y. PubMed DOI PMC

Ionic Environment Affects Biomolecular Interactions of Amyloid-β: SPR Biosensor Study