Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1995 Dec;69(6):2695–2702. doi: 10.1016/S0006-3495(95)80140-2

Spectroscopic studies of a phosphoinositide-binding peptide from gelsolin: behavior in solutions of mixed solvent and anionic micelles.

W Xian 1, R Vegners 1, P A Janmey 1, W H Braunlin 1
PMCID: PMC1236506  PMID: 8599675

Abstract

The peptide G(150-169) corresponds to a phosphatidylinositol 4,5-bisphosphate (PIP2) and filamentous actin (F-actin) binding site on gelsolin (residues 150-169, with the sequence KHVVPNEVVVQRLFQVKGRR). The conformation of this peptide in trifluoroethanol (TFE) aqueous solution was determined by 1H nuclear magnetic resonance as the first step toward understanding the structural aspects of the interaction of G(150-169) and PIP2. The circular dichroism experiments show that G(150-169) adopts a predominantly alpha-helical form in both 50% TFE aqueous solution and in the presence of PIP2 micelles, therefore establishing a connection between the two conformations. 1H nuclear magnetic resonance experiments of G(150-169) in TFE co-solvent show that the helical region extends from Pro-154 to Lys-166. The amphiphilic nature of this helical structure may be the key to understanding the binding of the peptide to lipids. Sodium dodecyl sulfate micelle solution is used as a model for anionic lipid environments. Preliminary studies of the conformation of G(150-169) in sodium dodecyl sulfate micelle solution show that the peptide forms an alpha-helix similar to but with some structural differences from that in TFE co-solvent. Fluorescence experiments provide evidence of peptide clustering over a narrow range of peptide/PIP2 ratios, which is potentially relevant to the biological function of PIP2.

Full text

PDF
2695

Images in this article

2700FIGURE 7
on p.2700

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brandts J. F., Halvorson H. R., Brennan M. Consideration of the Possibility that the slow step in protein denaturation reactions is due to cis-trans isomerism of proline residues. Biochemistry. 1975 Nov 4;14(22):4953–4963. doi: 10.1021/bi00693a026. [DOI] [PubMed] [Google Scholar]
  2. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  3. Geourjon C., Deléage G. SOPM: a self-optimized method for protein secondary structure prediction. Protein Eng. 1994 Feb;7(2):157–164. doi: 10.1093/protein/7.2.157. [DOI] [PubMed] [Google Scholar]
  4. Janmey P. A., Lamb J., Allen P. G., Matsudaira P. T. Phosphoinositide-binding peptides derived from the sequences of gelsolin and villin. J Biol Chem. 1992 Jun 15;267(17):11818–11823. [PubMed] [Google Scholar]
  5. Janmey P. A., Stossel T. P. Modulation of gelsolin function by phosphatidylinositol 4,5-bisphosphate. Nature. 1987 Jan 22;325(6102):362–364. doi: 10.1038/325362a0. [DOI] [PubMed] [Google Scholar]
  6. Janmey P. Cell biology. A slice of the actin. Nature. 1993 Aug 19;364(6439):675–676. doi: 10.1038/364675a0. [DOI] [PubMed] [Google Scholar]
  7. Lehrman S. R., Tuls J. L., Lund M. Peptide alpha-helicity in aqueous trifluoroethanol: correlations with predicted alpha-helicity and the secondary structure of the corresponding regions of bovine growth hormone. Biochemistry. 1990 Jun 12;29(23):5590–5596. doi: 10.1021/bi00475a025. [DOI] [PubMed] [Google Scholar]
  8. McLaughlin P. J., Gooch J. T., Mannherz H. G., Weeds A. G. Structure of gelsolin segment 1-actin complex and the mechanism of filament severing. Nature. 1993 Aug 19;364(6439):685–692. doi: 10.1038/364685a0. [DOI] [PubMed] [Google Scholar]
  9. Nelson J. W., Kallenbach N. R. Stabilization of the ribonuclease S-peptide alpha-helix by trifluoroethanol. Proteins. 1986 Nov;1(3):211–217. doi: 10.1002/prot.340010303. [DOI] [PubMed] [Google Scholar]
  10. Nishikawa K. Assessment of secondary-structure prediction of proteins. Comparison of computerized Chou-Fasman method with others. Biochim Biophys Acta. 1983 Oct 28;748(2):285–299. doi: 10.1016/0167-4838(83)90306-0. [DOI] [PubMed] [Google Scholar]
  11. Pasta P., Carrea G., Longhi R., Zetta L. Circular dichroism and proteolysis of human beta-endorphin in surfactant and lipid solutions. Biochim Biophys Acta. 1990 May 31;1039(1):1–4. doi: 10.1016/0167-4838(90)90218-5. [DOI] [PubMed] [Google Scholar]
  12. Ramachandran G. N., Mitra A. K. An explanation for the rare occurrence of cis peptide units in proteins and polypeptides. J Mol Biol. 1976 Oct 15;107(1):85–92. doi: 10.1016/s0022-2836(76)80019-8. [DOI] [PubMed] [Google Scholar]
  13. Richardson J. S., Richardson D. C. Amino acid preferences for specific locations at the ends of alpha helices. Science. 1988 Jun 17;240(4859):1648–1652. doi: 10.1126/science.3381086. [DOI] [PubMed] [Google Scholar]
  14. Sönnichsen F. D., Van Eyk J. E., Hodges R. S., Sykes B. D. Effect of trifluoroethanol on protein secondary structure: an NMR and CD study using a synthetic actin peptide. Biochemistry. 1992 Sep 22;31(37):8790–8798. doi: 10.1021/bi00152a015. [DOI] [PubMed] [Google Scholar]
  15. Wishart D. S., Sykes B. D., Richards F. M. Relationship between nuclear magnetic resonance chemical shift and protein secondary structure. J Mol Biol. 1991 Nov 20;222(2):311–333. doi: 10.1016/0022-2836(91)90214-q. [DOI] [PubMed] [Google Scholar]
  16. Wu C. S., Yang J. T. Sequence-dependent conformations of short polypeptides in a hydrophobic environment. Mol Cell Biochem. 1981 Oct 30;40(2):109–122. doi: 10.1007/BF00224754. [DOI] [PubMed] [Google Scholar]
  17. Yang J. T., Wu C. S., Martinez H. M. Calculation of protein conformation from circular dichroism. Methods Enzymol. 1986;130:208–269. doi: 10.1016/0076-6879(86)30013-2. [DOI] [PubMed] [Google Scholar]
  18. Yu F. X., Sun H. Q., Janmey P. A., Yin H. L. Identification of a polyphosphoinositide-binding sequence in an actin monomer-binding domain of gelsolin. J Biol Chem. 1992 Jul 25;267(21):14616–14621. [PubMed] [Google Scholar]
  19. Yun R. H., Anderson A., Hermans J. Proline in alpha-helix: stability and conformation studied by dynamics simulation. Proteins. 1991;10(3):219–228. doi: 10.1002/prot.340100306. [DOI] [PubMed] [Google Scholar]
  20. Zhou N. E., Kay C. M., Sykes B. D., Hodges R. S. A single-stranded amphipathic alpha-helix in aqueous solution: design, structural characterization, and its application for determining alpha-helical propensities of amino acids. Biochemistry. 1993 Jun 22;32(24):6190–6197. doi: 10.1021/bi00075a011. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES