Advertisement

Parasitology Research

, Volume 117, Issue 11, pp 3437–3446 | Cite as

In vitro interaction between Plasmodium falciparum myosin B (PfMyoB) and myosin A tail interacting protein (MTIP)

  • Paula C. Hernández
  • Moisés Wasserman
  • Jacqueline Chaparro-Olaya
Original Paper

Abstract

Apicomplexan parasites, including Plasmodium falciparum, are obligate intracellular organisms that utilize a strategy termed “gliding” to move and invade host cells, causing disease. Gliding is carried out by a protein complex known as the glideosome, which includes an actin–myosin motor. To date, six myosins have been identified in P. falciparum (PfMyoA, B, C, D, E, and F), but only the role of PfMyoA, the myosin of the glideosome that is involved in the process of red blood cell and mosquito cell invasion, has been established. Based on previous observations, we speculated that PfMyoA and PfMyoB may have similar or redundant functions. To test this hypothesis, we searched for in vitro interactions between PfMyoB and MTIP (myosin A tail interacting protein), the myosin light chain of PfMyoA. A set of differentially tagged PfMyoA, PfMyoB, and MTIP recombinant proteins was employed to specifically and simultaneously detect each myosin in competition assays and inhibition assays using specific peptides. MTIP potentially acts as the light chain of PfMyoB.

Keywords

Plasmodium falciparum Myosins Protein–protein interactions Pulldown Far western blot 

Notes

Acknowledgments

This work was supported by COLCIENCIAS (projects 110152128729 and 130834319109) and Universidad El Bosque (project UB-271-2010). Funding organizations had no role in study design, data analysis, decision to publish, or preparation of the manuscript. We want to thank Professors Celia R. Garcia and Chuck Farah of Universidade de Sao Paulo for their scientific and technical advice.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Bergman LW, Kaiser K, Fujioka H, Coppens I, Daly TM, Fox S et al (2003) Myosin a tail domain interacting protein (MTIP) localizes to the inner membrane complex of Plasmodium sporozoites. J Cell Sci 116:39–49CrossRefGoogle Scholar
  2. Bookwalter CS, Tay CL, McCrorie R, Previs MJ, Lu H, Krementsova EB, Fagnant PM, Baum J, Trybus KM (2017) Reconstitution of the core of the malaria parasite glideosome with recombinant Plasmodium class XIV myosin a and Plasmodium actin. J Biol Chem 292(47):19290–19303CrossRefGoogle Scholar
  3. Bosch J, Turley S, Daly TM, Bogh SM, Villasmil ML, Roach C, Zhou N, Morrisey JM, Vaidya AB, Bergman LW, Hol WGJ (2006) Structure of the MTIP-MyoA complex, a key component of the malaria parasite invasion motor. Proc Natl Acad Sci U S A 103:4852–4857CrossRefGoogle Scholar
  4. Bosch J, Turley S, Roach CM, Daly TM, Bergman LW, Hol WG (2007) The closed MTIP-myosin A-tail complex from the malaria parasite invasion machinery. J Mol Biol 372:77–88CrossRefGoogle Scholar
  5. Chaparro-Olaya J, Dluzewski AR, Margos G, Wasserman M, Mitchell GH, Bannister L et al (2003) The multiple myosins of malaria: the smallest malaria myosin, Plasmodium falciparum myosin-B (Pfmyo-B) is expressed in mature schizonts and merozoites. Eur J Protistol 39(4):423–427CrossRefGoogle Scholar
  6. Douse CH, Green JL, Salgado PS, Simpson PJ, Thomas JC, Langsley G, Holder AA, Tate EW, Cota E (2012) Regulation of the Plasmodium motor complex: phosphorylation of myosin a tail-interacting protein (MTIP) loosens its grip on MyoA. J Biol Chem 287(44):36968–36977CrossRefGoogle Scholar
  7. Douse CH, Vrielink N, Wenlin Z, Cota E, Tate EW (2015) Targeting a dynamic protein–protein interaction: fragment screening against the malaria myosin a motor complex. ChemMedChem 10(1):134–143CrossRefGoogle Scholar
  8. Einarson MB, Pugacheva EN, Orlinick JR (2007) GST Pull-down. CSH Protoc 2007:pdb.prot4757PubMedGoogle Scholar
  9. Farrow RE, Green J, Katsimitsoulia Z, Taylor WR, Holder AA, Molloy JE (2011) The mechanism of erythrocyte invasion by the malarial parasite, Plasmodium falciparum. Semin Cell Dev Biol 22(9):953–960CrossRefGoogle Scholar
  10. Green JL, Martin SR, Fielden J, Ksagoni A, Grainger M, Yim Lim BY et al (2006) The MTIP–myosin A complex in blood stage malaria parasites. J Mol Biol 355(5):933–941CrossRefGoogle Scholar
  11. Green JL, Wall RJ, Vahokoski J, Yusuf NA, Ridzuan MAM, Stanway RR, Stock J, Knuepfer E, Brady D, Martin SR, Howell SA, Pires IP, Moon RW, Molloy JE, Kursula I, Tewari R, Holder AA (2017) Compositional and expression analyses of the glideosome during the Plasmodium life cycle reveal an additional myosin light chain required for maximum motility. J Biol Chem 292(43):17857–17875CrossRefGoogle Scholar
  12. Guttery DS, Poulin B, Ferguson DJ, Szöőr B, Wickstead B, Carroll PL et al (2012) A unique protein phosphatase with kelch-like domains (PPKL) in Plasmodium modulates ookinete differentiation, motility and invasion. PLoS Pathog 8(9):e1002948CrossRefGoogle Scholar
  13. Hernández PC, Morales L, Castellanos IC, Wasserman M, Chaparro-Olaya J (2017) Myosin B of Plasmodium falciparum (PfMyoB): in silico prediction of its three-dimensional structure and its possible interaction with MTIP. Parasitol Res 116(4):1373–1382CrossRefGoogle Scholar
  14. Jones ML, Kitson EL, Rayner JC (2006) Plasmodium falciparum erythrocyte invasion: a conserved myosin associated complex. Mol Biochem Parasitol 147(1):74–84CrossRefGoogle Scholar
  15. Koch M, Baum J (2016) The mechanics of malaria parasite invasion of the human erythrocyte—towards a reassessment of the host cell contribution. Cell Microbiol 18(3):319–329CrossRefGoogle Scholar
  16. Kortagere S, Welsh WJ, Morrisey JM, Daly T, Ejigiri I, Sinnis P, Vaidya AB, Bergman LW (2010) Structure-based design of novel small-molecule inhibitors of Plasmodium falciparum. J Chem Inf Model 50(5):840–849CrossRefGoogle Scholar
  17. Patzewitz EM, Guttery DS, Poulin B, Ramakrishnan C, Ferguson DJ, Wall RJ et al (2013) An ancient protein phosphatase, SHLP1, is critical to microneme development in Plasmodium ookinetes and parasite transmission. Cell Rep 3(3):622–629CrossRefGoogle Scholar
  18. Otto TD, Wilinski D, Assefa S, Keane TM, Sarry LR, Böhme U, Lemieux J, Barrell B, Pain A, Berriman M, Newbold C, Llinás M (2010) New insights into the blood stage transcriptome of Plasmodium falciparum using RNA-Seq. Mol Microbiol 76(1):12–24CrossRefGoogle Scholar
  19. Rivadeneira EM, Wasserman M, Espinal CT (1983) Separation and concentration of schizonts of Plasmodium falciparum by Percoll gradients. J Protozool 30(2):367–370CrossRefGoogle Scholar
  20. Sambrook J, Russell DW (2006) Detection of protein-protein interactions using the GST fusion protein pulldown technique. CSH Protoc 2006(1)Google Scholar
  21. Tardieux I, Baum J (2016) Reassessing the mechanics of parasite motility and host-cell invasion. J Cell Biol 214:507–515CrossRefGoogle Scholar
  22. Thomas JC, Green JL, Howson RI, Simpson P, Moss DK, Martin SR, Holder AA, Cota E, Tate EW (2010) Interaction and dynamics of the Plasmodium falciparum MTIP–MyoA complex, a key component of the invasion motor in the malaria parasite. Mol BioSyst 6:494–498CrossRefGoogle Scholar
  23. WHO (2017) World malaria report 2017. http://www.who.int/malaria/publications/world-malaria-report-2017/en/. Accessed 01 May 2018
  24. Williamson RC, Bass MD (2015) Comparing the affinity of GTPase-binding proteins using competition assays. J Vis Exp 104Google Scholar
  25. Wu Y, Li Q, Chen XZ (2007) Detecting protein–protein interactions by far western blotting. Nat Protoc 2(12):3278–3284CrossRefGoogle Scholar
  26. Yusuf NA, Green JL, Wall RJ, Knuepfer E, Moon RW, Schulte-Huxel C, Stanway RR, Martin SR, Howell SA, Douse CH, Cota E, Tate EW, Tewari R, Holder AA (2015) The Plasmodium class XIV myosin, MyoB, has a distinct subcellular location in invasive and motile stages of the malaria parasite and an unusual light chain. J Biol Chem 290(19):12147–12164CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratorio de Parasitología Molecular Instituto de Biología MolecularUniversidad El BosqueBogotáColombia
  2. 2.Laboratorio de Investigaciones Básicas en BioquímicaUniversidad Nacional de ColombiaBogotáColombia

Personalised recommendations