Advertisement

Cell and Tissue Biology

, Volume 11, Issue 3, pp 227–233 | Cite as

The relationship between micelle formation and biological activity of peptide 562–572 of luteinizing hormone receptor modified with decanoyl radicals

  • E. A. Shpakova
  • V. N. Sorokoumov
  • A. V. Akent’ev
  • K. V. Derkach
  • T. B. Tennikova
  • A. O. ShpakovEmail author
Article
  • 17 Downloads

Abstract

Lipophilic derivatives of peptides corresponding to cytoplasmic regions of G protein-coupled receptors (GPCR) can act as intracellular agonists. Our previous work showed that peptides corresponding to residues 562–572 of luteinizing hormone receptor and modified with palmitate or decanoate at the C-terminus activate adenylate cyclase in rat testes. The stimulating effect of peptide 562–572 modified with decanoates at both the N- and the C-termini (peptide IV) reached its maximum at the peptide concentration of 10–5 M and diminished with further increase in its concentration. It was supposed that this effect was due to peptide IV ability to form micelles. To verify this hypothesis, the relationship between biological activity, hydrophobicity, and ability to form micelles was investigated for peptide IV and other acylated derivatives of peptide 562–572, including those carrying C-terminal decanoate (peptide III) and palmitate (peptide VI) moieties. It was found that the stimulating effect of peptide IV taken in the concentration of 10–5 M on adenylate cyclase activity in plasma membranes of rat testes and ovaries was only slightly lower than that of peptide VI and higher than the effect of peptide III. At the concentration of 10–3 M, the effect of peptide IV was 20–27% lower and amounted to only 50–51 and 87–88% of the effects of peptides VI and III, respectively. In spite of its high hydrophobicity, peptide IV was characterized with an abnormally low retention time when eluted from a Nucleosil C8 column during reverse-phase HPLC: it was even lower than the retention time of nonmodified peptide 562–572. However, the retention time of peptide IV, but not of other peptides, increased significantly when the eluent contained a higher proportion of trifluoroacetic acid, which disrupts micellelike structures (0.5 instead of 0.1%). The surface tension of peptide IV solution in water slightly decreased with increasing peptide concentration, but rapidly dropped and reached a plateau at the concentration of 7 × 10–6 M, which indicates the beginning of micelle formation. Thus, peptide IV in the concentrations above 10–5 M forms micelles, which prevents it from interacting with the receptor. The ability of GPCR peptides to aggregate and form micelles should be taken into account in the development of their new membrane-active analogs.

Keywords

peptide luteinizing hormone receptor hydrophobicity micelle formation adenylate cyclase decanoyl radical 

Abbreviations

AC

adenylate cyclase

ACSS

adenylyl cyclase sygnaling system

CCM

critical concentration of micelle formation

LHR

luteinizing hormone receptor

TFAA

trifluoroacetic acid

GPCR

G protein-coupled receptor

Dec

decanoic acid residue

Palm

palmitic acid residue

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Avrahami, D. and Shai, Y., A new group of antifungal and antibacterial lipopeptides derived from non-membrane active peptides conjugated to palmitic acid, J. Biol. Chem., 2004, vol. 279, pp. 12277–12285.CrossRefPubMedGoogle Scholar
  2. Chopineau, J., Robert, S., Fénart, L., Cecchelli, R., Lagoutte, B., Paitier, S., Dehouck, M.P., and Domurado, D., Monoacylation of ribonuclease A Enables its transport across an in vitro model of the blood–brain barrier, J. Control Release, 1998, vol. 56, pp. 231–237.CrossRefPubMedGoogle Scholar
  3. Cisowski, J., O’Callaghan, K., Kuliopulos, A., Yang, J., Nguyen, N., Deng, Q., Yang, E., Fogel, M., Tressel, S., Foley, C., Agarwal, A., Hunt, S.W., 3rd, McMurry, T., Brinckerhoff, L., and Covic, L., Targeting protease-activated receptor-1 with cell-penetrating pepducins in lung cancer, Am. J. Pathol., 2011, vol. 179, pp. 513–523.CrossRefPubMedPubMedCentralGoogle Scholar
  4. Covic, L., Gresser, A.L., Talavera, J., Swift, S., and Kuliopulos, A., Activation and inhibition of G protein-coupled receptors by cell-penetrating membrane-tethered peptides, Proc. Natl. Acad. Sci. U. S. A., 2002, vol. 99, pp. 643–648.CrossRefPubMedPubMedCentralGoogle Scholar
  5. Derkach, K.V., Shpakova, E.A., and Shpakov, A.O., Palmitoylated peptide 562–572 of luteinizing hormone receptor increases testosterone level in male rats, Bull. Exp. Biol. Med., 2014, vol. 158, no. 2, pp. 209–212.CrossRefPubMedGoogle Scholar
  6. Derkach, K.V., Shpakova, E.A., Titov, A.M., and Shpakov, A.O., Intranasal and intramuscular administration of lysine-palmitoylated peptide 612–627 of thyroidstimulating hormone receptor increases the level of thyroid hormones in rats, Int. J. Pept. Res. Ther., 2015, vol. 21, pp. 249–260.CrossRefGoogle Scholar
  7. Geng, X. and Regnier, F.E., Retention model for proteins in reversed-phase liquid chromatography, J. Chromatogr., 1984, vol. 296, pp. 15–30.CrossRefPubMedGoogle Scholar
  8. Mas-Moruno, C., Cascales, L., Cruz, L.J., Mora, P., Pérez-Payá, E., and Albericio, F., Nanostructure formation enhances the activity of LPS-neutralizing peptides, ChemMedChem, 2008, vol. 3, pp. 1748–1755.CrossRefPubMedGoogle Scholar
  9. Naeem, A., Ashraf, M.T., Akram, M., and Khan, R.H., Comparative study of effects of polyols, salts, and alcohols on trichloroacetic acid-induced state of cytochrome c, Biochemistry (Moscow), 2006, vol. 71, pp. 1101–1109.CrossRefGoogle Scholar
  10. Severino, B., Incisivo, G.M., Fiorino, F., Bertolino, A., Frecentese, F., Barbato, F., Manganelli, S., Maggioni, G., Capasso, D., Caliendo, G., Santagada, V., Sorrentino, R., Roviezzo, F., and Perissutti, E., Identification of a pepducin acting as S1P3 receptor antagonist, J. Pept. Sci., 2013, vol. 19, pp. 717–724.CrossRefPubMedGoogle Scholar
  11. Shpakov, A.O., Signal protein-derived peptides as functional probes and regulators of intracellular signaling, J. Amino Acids, 2011. doi 10.4061/2011/656051Google Scholar
  12. Shpakov, A.O. and Derkach, K.V., New achievements in development and application of GPCR-peptides, J. Evol. Biochem. Physiol., 2015, vol. 51, no. 1, pp. 11–18.CrossRefGoogle Scholar
  13. Shpakov, A.O., and Pertseva, M.N., The peptide strategy as a novel approach to the study of G protein-coupled signaling systems, in Signal Transduction Research Trends, Grachevsky, N.O., Ed., Nova Science Publishers, Inc., 2007, pp. 45–93.Google Scholar
  14. Shpakov, A.O. and Shpakova, E.A., The prospects for use of peptides and their derivatives, structurally corresponding to the G protein-coupled receptors, in medicine, Biochemistry (Moscow), Suppl. Ser. B: Biomed. Chem., 2014a, vol. 8, pp. 19–26.CrossRefGoogle Scholar
  15. Shpakov, A.O. and Shpakova, E.A., The use of peptides derived from G protein-coupled receptors and heterotrimeric G proteins in the study of their structure and functions (chapter 4), in Protein Purification and Analysis. III. Methods and Applications, Concept Press Ltd., 2014b. http://www.iconceptpress.com/books/protein-purificationand-analysis-iii–methods-and-applications/.Google Scholar
  16. Shpakov, A.O., Gur’yanov, I.A., Kuznetsova, L.A., Plesneva, S.A., Shpakova, E.A., Vlasov, G.P., and Pertseva, M.N., Studies of the molecular mechanisms of action of relaxin on the adenylyl cyclase signaling system using synthetic peptides derived from the LGR7 relaxin receptor, Neurosci. Behav. Physiol., 2007, vol. 37, pp. 705–714.CrossRefPubMedGoogle Scholar
  17. Shpakov, A.O., Derkach, K.V., and Bondareva, V.M., A decrease of the sensitivity of adenylyl cyclase and heterotrimeric G proteins to chorionic gonadotrophin and peptide hormones action in the tissues of reproductive system in rats with experimental type 2 diabetes, Biochemistry (Moscow), Suppl. Ser. B: Biomed. Chem., 2010a, vol. 4, no. 3, pp. 258–263.CrossRefGoogle Scholar
  18. Shpakov, A.O., Shpakova, E.A., Tarasenko, I.I., Derkach, K.V., and Vlasov, G.P., The peptides mimicking the third intracellular loop of 5-hydroxytryptamine receptors of the types 1B and 6 selectively activate G proteins and receptor-specifically inhibit serotonin signaling via the adenylyl cyclase system, Int. J. Pept. Res. Ther., 2010b, vol. 16, pp. 95–105.CrossRefGoogle Scholar
  19. Shpakov, A.O., Shpakova, E.A., Tarasenko, I.I., and Derkach, K.V., Receptor and tissue specificity of the effect of peptides corresponding to intracellular regions of the serpentine type receptors, Biochemistry (Moscow), Suppl. Ser. A: Membr. Cell Biol., 2011, vol. 6, no. 1, pp. 16–25.CrossRefGoogle Scholar
  20. Shpakov, A.O., Chistyakova, O.V., Derkach, K.V., Moiseyuk, I.V., and Bondareva, V.M., Intranasal insulin affects adenylyl cyclase system in rat tissues in neonatal diabetes, Central Eur. J. Biol., 2012a, vol. 7, pp. 33–47.Google Scholar
  21. Shpakov, A.O., Shpakova, E.A., and Derkach, K.V., The sensitivity of the adenylyl cyclase system in rat thyroidal and extrathyroidal tissues to peptides corresponding to the third intracellular loop of thyroid-stimulating hormone receptor, Curr. Top. Pept. Prot. Res., 2012b, vol. 13, pp. 61–73.Google Scholar
  22. Shpakova, E.A. and Shpakov, A.O., Regulation of adenylyl cyclase activity in rat testes by acylated derivatives of peptide 562–572 of a luteinizing hormone receptor, Cell Tissue Biol., 2013, vol. 8, no. 2, pp. 152–159.CrossRefGoogle Scholar
  23. Shpakova, E.A., Derkach, K.V., and Shpakov, A.O., Biological activity of lipophilic derivatives of peptide 562–572 of rat luteinizing hormone receptor, Dokl. Biochem. Biophys., 2013, vol. 452, no. 1, pp. 248–250.CrossRefPubMedGoogle Scholar
  24. Sikorska, E. and Kwiatkowska, A., Micelle-bound conformations of neurohypophyseal hormone analogues modified with a Cα-disubstituted residue: NMR and molecular modelling studies, J. Biomol. Struct. Dyn., 2013, vol. 31, pp. 748–764.CrossRefPubMedGoogle Scholar
  25. Tressel, S.L., Koukos, G., Tchernychev, B., Jacques, S.L., Covic, L., and Kuliopulos, A., pharmacology, biodistribution, and efficacy of GPCR-based pepducins in disease models, Meth. Mol. Biol., 2011, vol. 683, pp. 259–275.CrossRefGoogle Scholar
  26. Vad, B., Thomsen, L.A., Bertelsen, K., Franzmann, M., Pedersen, J.M., Nielsen, S.B., Vosegaard, T., Valnickova, Z., Skrydstrup, T., Enghild, J.J., Wimmer, R., Nielsen, N.C., and Otzen, D.E., Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide, Biochim. Biophys. Acta, 2010, vol. 1804, pp. 806–820.CrossRefPubMedGoogle Scholar
  27. Winkel, D., Theoretical refinement of the pendant drop method for measuring surface tensions, J. Phys. Chem., 1965, vol. 69, pp. 348–350.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • E. A. Shpakova
    • 1
  • V. N. Sorokoumov
    • 1
  • A. V. Akent’ev
    • 1
  • K. V. Derkach
    • 2
  • T. B. Tennikova
    • 1
  • A. O. Shpakov
    • 2
    Email author
  1. 1.Institute of ChemistrySt. Petersburg State UniversitySt. PetersburgRussia
  2. 2.Sechenov Institute of Evolutionary Biology and BiochemistryRussian Academy of SciencesSt. PetersburgRussia

Personalised recommendations