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Metabolic Brain Disease

, Volume 33, Issue 6, pp 1975–1984 | Cite as

Changes in biophysical characteristics of PFN1 due to mutation causing amyotrophic lateral sclerosis

  • Mina Nekouei
  • Parviz Ghezellou
  • Atousa Aliahmadi
  • Sareh Arjmand
  • Mahmoud Kiaei
  • Alireza Ghassempour
Original Article

Abstract

Single amino acid mutations in profilin 1 (PFN1) have been found to cause amyotrophic lateral sclerosis (ALS). Recently, we developed a mouse model for ALS using a PFN1 mutation (glycine 118 to valine, G118V), and we are now interested in understanding how PFN1 becomes toxically lethal with only one amino acid substitution. Therefore, we studied mutation-related changes in the PFN1 protein and hypothesized that such changes significantly disturb its structure. Initially, we expressed and studied the purified PFN1WT and PFN1G118V proteins from bacterial culture. We found that the PFN1G118V protein has a different mean residue ellipticity, as measured by far-UV circular dichroism, accompanied by a spectral shift. The intrinsic fluorescence of PFN1G118V showed a small fluctuation in maximum fluorescence absorption and intensity. Moreover, we examined the time course of PFN1 aggregation using SDS-PAGE, western blotting, and MALDI-TOF/TOF and found that compared with PFN1WT, PFN1G118V had an increased tendency to form aggregates. Dynamic light scattering data confirmed this, showing a larger size distribution for PFN1G118V. Our data explain why PFN1G118V tends to aggregate, a phenotype that may be the basis for its neurotoxicity.

Keywords

PFN1G118V PFN1WT ALS Aggregation Actin binding domain 

Notes

Acknowledgements

We gratefully acknowledge Dr. Seyed Omid Ranaei Siadat from the Protein Research Center (Shahid Beheshti University) for allowing us to access his laboratory, which focuses on protein expression. Financial support by the Shahid Beheshti University Research Council is also gratefully acknowledged. This manuscript was edited by the Science Communication Group at the University of Arkansas for Medical Sciences (UAMS); we also thank Caroline Barham (UAMS) for proofreading the manuscript. The authors acknowledge funding from the UAMS startup fund, UAMS Center for Translational Neurosciences, NIGMS IDeA Program Award P30 GM110702, P20GM109005, and NINDS NS088653 and NS101334.

Author contributions

M.K. introduced the idea, designed the study, analyzed the data, and wrote the manuscript. A.G. designed the study, analyzed the data, and wrote the manuscript. M.N. designed and performed experiments, analyzed data, and wrote the manuscript. P.G. assisted with MALDI analysis and helped prepare the manuscript. A.A. assisted with the expression of recombinant PFN1, and S.A. assisted with sub-cloning experiments.

Compliance with ethical standards

Conflict of interest

Dr. Kiaei and UAMS have a financial interest in the technology discussed in this publication. These financial interests have been reviewed and approved in accordance with the UAMS conflict of interest policies. Other authors have no conflict to disclose.

Supplementary material

11011_2018_305_MOESM1_ESM.docx (1.2 mb)
ESM 1 (DOCX 1242 kb)

References

  1. Alkam D, Feldman EZ, Singh A, Kiaei M (2017) Profilin1 biology and its mutation, actin (g) in disease. Cell Mol Life Sci 74:967–981CrossRefGoogle Scholar
  2. Boopathy S, Silvas TV, Tischbein M, Jansen S, Shandilya SM, Zitzewitz JA, Landers JE, Goode BL, Schiffer CA, Bosco DA (2015) Structural basis for mutation-induced destabilization of profilin 1 in ALS. Proc Natl Acad Sci USA 112:7984–7989CrossRefGoogle Scholar
  3. Chen Y, Zheng ZZ, Huang R, Chen K, Song W, Zhao B, Chen X, Yang Y, Yuan L, Shang HF (2013) PFN1 mutations are rare in Han Chinese populations with amyotrophic lateral sclerosis. Neurobiol Aging 34(7):1922 e1–e5CrossRefGoogle Scholar
  4. Del Poggetto E, Bemporad F, Tatini F, Chiti F (2015a) Mutations of Profilin-1 associated with amyotrophic lateral sclerosis promote aggregation due to structural changes of its native state. ACS Chem Biol 10:2553–2563CrossRefGoogle Scholar
  5. Del Poggetto E, Chiti F, Bemporad F (2015b) The folding process of human Profilin-1, a novel protein associated with familial amyotrophic lateral sclerosis. Sci Report 5:–12332Google Scholar
  6. Del Poggetto E, Gori L, Chiti F (2016) Biophysical analysis of three novel profilin-1 variants associated with amyotrophic lateral sclerosis indicates a correlation between their aggregation propensity and the structural features of their globular state. Biol Chem 397:927–937PubMedGoogle Scholar
  7. Fan L, Simard LR (2002) Survival motor neuron (SMN) protein: role in neurite outgrowth and neuromuscular maturation during neuronal differentiation and development. Hum Mol Genet 11:1605–1614CrossRefGoogle Scholar
  8. Fedorov A, Pollard T, Almo S (1994) Purification, characterization and crystallization of human platelet profilin expressed in Escherichia coli. J Mol Biol 241:480–482CrossRefGoogle Scholar
  9. Fil D, DeLoach A, Yadav S, Alkam D, MacNicol M, Singh A, Compadre CM, Goellner JJ, O'Brien CA, Fahmi T, Basnakian AG, Calingasan NY, Klessner JL, Beal FM, Peters OM, Metterville J, Brown RH Jr, Ling KKY, Rigo F, Ozdinler PH, Kiaei M (2017) Mutant Profilin1 transgenic mice recapitulate cardinal features of motor neuron disease. Hum Mol Genet 26:686–701PubMedGoogle Scholar
  10. Freischmidt A, Schopflin M, Feiler MS, Fleck AK, Ludolph AC, Weishaupt JH (2015) Profilin 1 with the amyotrophic lateral sclerosis associated mutation T109M displays unaltered actin binding and does not affect the actin cytoskeleton. BMC Neurosci 16:77CrossRefGoogle Scholar
  11. Hardiman O, Al-Chalabi A, Chio A, Corr EM, Logroscino G, Robberecht W, Shaw PJ, Simmons Z, van den Berg LH (2017) Amyotrophic lateral sclerosis. Nat Rev Dis Primers 3:17071CrossRefGoogle Scholar
  12. Hensel N, Claus P (2018) The actin cytoskeleton in SMA and ALS: how does it contribute to Motoneuron degeneration? Neuroscientist 24(1):54–72CrossRefGoogle Scholar
  13. Ingre C, Landers JE, Rizik N, Volk AE, Akimoto C, Birve A, Hübers A, Keagle PJ, Piotrowska K, Press R, Andersen PM, Ludolph AC, Weishaupt JHIngre C, Landers JE, Rizik N, Volk AE, Akimoto C, Birve A, Hübers A, Keagle PJ, Piotrowska K, Press R, Andersen PM, Ludolph AC, Weishaupt JH (2013) A novel phosphorylation site mutation in profilin 1 revealed in a large screen of US, Nordic, and German amyotrophic lateral sclerosis/frontotemporal dementia cohorts. Neurobiol Aging 34(6):1708 e1–e6CrossRefGoogle Scholar
  14. Johnson JO, Mandrioli J, Benatar M, Abramzon Y, van Deerlin V, Trojanowski JQ, Gibbs JR, Brunetti M, Gronka S, Wuu J, Ding J, McCluskey L, Martinez-Lage M, Falcone D, Hernandez DG, Arepalli S, Chong S, Schymick JC, Rothstein J, Landi F, Wang YD, Calvo A, Mora G, Sabatelli M, Monsurrò MR, Battistini S, Salvi F, Spataro R, Sola P, Borghero G, ITALSGEN Consortium, Galassi G, Scholz SW, Taylor JP, Restagno G, Chiò A, Traynor BJ (2010) Exome sequencing reveals VCP mutations as a cause of familial ALS. Neuron 68:857–864CrossRefGoogle Scholar
  15. Kohn J, Wilchek M (1984) The use of cyanogen bromide and other novel cyanylating agents for the activation of polysaccharide resins. Appl. Biochem. Biotechnol. 9:285–305CrossRefGoogle Scholar
  16. Krishnan K, Holub O, Gratton E, Clayton AH, Cody S, Moens PD (2009) Profilin interaction with phosphatidylinositol (4,5)-bisphosphate destabilizes the membrane of giant unilamellar vesicles. Biophys J 96:5112–5121CrossRefGoogle Scholar
  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  18. Lambrechts A, Jonckheere V, Dewitte D, Vandekerckhove J, Ampe C (2002) Mutational analysis of human profilin I reveals a second PI(4,5)-P2 binding site neighbouring the poly (L-proline) binding site. BMC Biochem 3:12CrossRefGoogle Scholar
  19. Lim L, Kang J, Song J (2017) ALS-causing profilin-1-mutant forms a non-native helical structure in membrane environments. Biochim Biophys Acta 1859(11):2161–2170CrossRefGoogle Scholar
  20. Lorber B, Fischer F, Bailly M, Roy H, Kern D (2012) Protein analysis by dynamic light scattering: methods and techniques for students. Biochem Mol Biol Educ 40:372–382CrossRefGoogle Scholar
  21. Matsukawa K, Hashimoto T, Matsumoto T, Ihara R, Chihara T, Miura M, Wakabayashi T, Iwatsubo T (2016) Familial amyotrophic lateral sclerosis-linked mutations in profilin 1 exacerbate TDP-43-induced degeneration in the retina of drosophila melanogaster through an increase in the cytoplasmic localization of TDP-43. J Biol Chem 291:23464–23476CrossRefGoogle Scholar
  22. McLachlan GD, Cahill SM, Girvin ME, Almo SC (2007) Acid-induced equilibrium folding intermediate of human platelet profilin. Biochemistry 46:6931–6943CrossRefGoogle Scholar
  23. Metzler WJ, Farmer BT, Constantine KL, Friedrichs MS, Mueller L, Lavoie T (1995) Refined solution structure of human profilin I. Protein Sci 4:450–459CrossRefGoogle Scholar
  24. Nölle A, Zeug A, van Bergeijk J, Tönges L, Gerhard R, Brinkmann H, Al Rayes S, Hensel N, Schill Y, Apkhazava D, Jablonka S, O'mer J, Srivastav RK, Baasner A, Lingor P, Wirth B, Ponimaskin E, Niedenthal R, Grothe C, Claus P (2011) The spinal muscular atrophy disease protein SMN is linked to the Rho-kinase pathway via profilin. Hum Mol Genet 20(24):4865–4878CrossRefGoogle Scholar
  25. Schutt CE, Myslik JC, Rozycki MD, Goonesekere NC, Lindberg U (1993) The structure of crystalline profilin-beta-actin. Nature 365:810–816CrossRefGoogle Scholar
  26. Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M (2007) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860CrossRefGoogle Scholar
  27. Smith BN, Vance C, Scotter EL, Troakes C, Wong CH, Topp S, Maekawa S, King A, Mitchell JC, Lund K, Al-Chalabi A, Ticozzi N, Silani V, Sapp P, Brown Jr. RH, Landers JE, Al-Sarraj S, Shaw CE (2014) Novel mutations support a role for Profilin 1 in the pathogenesis of ALS. Neurobiol Aging 36:1602.e1617–1602.e1627Google Scholar
  28. Tanaka Y, Hasegawa M (2016) Profilin 1 mutants form aggregates that induce accumulation of prion-like TDP-43. Prion 10:283–289CrossRefGoogle Scholar
  29. Tanaka Y, Nonaka T, Suzuki G, Kametani F, Hasegawa M (2016) Gain-of-function profilin 1 mutations linked to familial amyotrophic lateral sclerosis cause seed-dependent intracellular TDP-43 aggregation. Hum Mol Genet 25:1420–1433CrossRefGoogle Scholar
  30. van Es MA, Hardiman O, Chio A, Al-Chalabi A, Pasterkamp RJ, Veldink JH, van den Berg LH (2017) Amyotrophic lateral sclerosis. Lancet 390(10107):2084-2098CrossRefGoogle Scholar
  31. Witke W (2004) The role of profilin complexes in cell motility and other cellular processes. Trends Cell Biol 14:461–469CrossRefGoogle Scholar
  32. Wu CH, Fallini C, Ticozzi N, Keagle PJ, Sapp PC, Piotrowska K, Lowe P, Koppers M, McKenna-Yasek D, Baron DM, Kost JE, Gonzalez-Perez P, Fox AD, Adams J, Taroni F, Tiloca C, Leclerc AL, Chafe SC, Mangroo D, Moore MJ, Zitzewitz JA, Xu ZS, van den Berg LH, Glass JD, Siciliano G, Cirulli ET, Goldstein DB, Salachas F, Meininger V, Rossoll W, Ratti A, Gellera C, Bosco DA, Bassell GJ, Silani V, Drory VE, Brown Jr RH, Landers JE (2012) Mutations in the profilin 1 gene cause familial amyotrophic lateral sclerosis. Nature 488:499–503CrossRefGoogle Scholar
  33. Yang C, Danielson EW, Qiao T, Metterville J, Brown RH Jr, Landers JE, Xu Z (2016) Mutant PFN1 causes ALS phenotypes and progressive motor neuron degeneration in mice by a gain of toxicity. Proc Natl Acad Sci U S A 113:E6209–E6218CrossRefGoogle Scholar
  34. Zarei S, Carr K, Reiley L, Diaz K, Guerra O, Altamirano OF, Pagani W, Lodin D, Orozco G, Chinea A (2015) A comprehensive review of amyotrophic lateral sclerosis. Surg Neurol Int 6:171CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Phytochemistry, Medicinal Plants and Drugs Research InstituteShahid Beheshti UniversityTehranIran
  2. 2.Department of Biology, Medicinal Plants and Drugs Research InstituteShahid Beheshti UniversityTehranIran
  3. 3.Protein Research CenterShahid Beheshti UniversityTehranIran
  4. 4.Department of Pharmacology and Toxicology, Department of Neurology, Department of GeriatricsUniversity of Arkansas for Medical SciencesLittle RockUSA

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