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Wavelength-Selective Fluorescence of a Model Transmembrane Peptide: Constrained Dynamics of Interfacial Tryptophan Anchors

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Abstract

WALPs are prototypical, α-helical transmembrane peptides that represent a consensus sequence for transmembrane segments of integral membrane proteins and serve as excellent models for exploring peptide-lipid interactions and hydrophobic mismatch in membranes. Importantly, the WALP peptides are in direct contact with the lipids. They consist of a central stretch of alternating hydrophobic alanine and leucine residues capped at both ends by tryptophans. In this work, we employ wavelength-selective fluorescence approaches to explore the intrinsic fluorescence of tryptophan residues in WALP23 in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes. Our results show that the four tryptophan residues in WALP23 exhibit an average red edge excitation shift (REES) of 6 nm, implying their localization at the membrane interface, characterized by a restricted microenvironment. This result is supported by fluorescence anisotropy and lifetime measurements as a function of wavelength displayed by WALP23 tryptophans in POPC membranes. These results provide a new approach based on intrinsic fluorescence of interfacial tryptophans to address protein-lipid interaction and hydrophobic mismatch.

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References

  1. Killian JA (2003) Synthetic peptides as models for intrinsic membrane proteins. FEBS Lett 555:134–138

    Article  CAS  Google Scholar 

  2. Killian JA, Salemink I, de Planque MRR, Lindblom G, Koeppe II RE, Greathouse DV (1996) Induction of nonbilayer structures in diacylphosphatidylcholine model membranes by transmembrane α-helical peptides: importance of hydrophobic mismatch and proposed role of tryptophans. Biochemistry 35:1037–1045

    Article  CAS  Google Scholar 

  3. Vostrikov VV, Koeppe II RE (2011) Response of GWALP transmembrane peptides to changes in the tryptophan anchor positions. Biochemistry 50:7522–7535

    Article  CAS  Google Scholar 

  4. Vostrikov VV, Daily AE, Greathouse DV, Koeppe II RE (2010) Charged or aromatic anchor residue dependence of transmembrane peptide tilt. J Biol Chem 285:31723–31730

    Article  CAS  Google Scholar 

  5. van’t Hag L, Li X, Meikle TG, Hoffmann SV, Jones NC, Pedersen JS, Hawley AM, Gras SL, Conn CE, Drummond CJ (2016) How peptide molecular structure and charge influence the nanostructure of lipid bicontinuous cubic mesophases: model synthetic WALP peptides provide insights. Langmuir 32:6882–6894

    Article  Google Scholar 

  6. Killian JA (1992) Gramicidin and gramicidin-lipid interactions. Biochim Biophys Acta 1113:391–425

    Article  CAS  Google Scholar 

  7. Koeppe II RE, Andersen OS (1996) Engineering the gramicidin channel. Annu Rev Biophys Biomol Struct 25:231–258

    Article  CAS  Google Scholar 

  8. Kelkar DA, Chattopadhyay A (2007) The gramicidin ion channel: a model membrane protein. Biochim Biophys Acta 1768:2011–2025

    Article  CAS  Google Scholar 

  9. Arkin IT, Brunger AT (1998) Statistical analysis of predicted transmembrane α-helices. Biochim Biophys Acta 1429:113–128

    Article  CAS  Google Scholar 

  10. Adamian L, Nanda V, DeGrado WF, Liang J (2005) Empirical lipid propensities of amino acid residues in multispan alpha helical membrane proteins. Proteins 59:496–509

    Article  CAS  Google Scholar 

  11. Strandberg E, Ӧzdirekcan S, Rijkers DTS, van der Wel PCA, Koeppe II RE, Liskamp RMJ, Killian JA (2004) Tilt angles of transmembrane model peptides in oriented and non-oriented lipid bilayers as determined by 2H solid-state NMR. Biophys J 86:3709–3721

    Article  CAS  Google Scholar 

  12. van der Wel PCA, Strandberg E, Killian JA, Koeppe II RE (2002) Geometry and intrinsic tilt of a tryptophan-anchored transmembrane alpha-helix determined by 2H NMR. Biophys J 83:1479–1488

  13. Schiffer M, Chang C-H, Stevens FJ (1992) The functions of tryptophan residues in membrane proteins. Protein Eng 5:213–214

    Article  CAS  Google Scholar 

  14. Kelkar DA, Chattopadhyay A (2006) Membrane interfacial localization of aromatic amino acids and membrane protein function. J Biosci 31:297–302

    Article  CAS  Google Scholar 

  15. Mouritsen OG, Bloom M (1984) Mattress model of lipid-protein interactions in membranes. Biophys J 46:141–153

    Article  CAS  Google Scholar 

  16. Killian JA (1998) Hydrophobic mismatch between proteins and lipids in membranes. Biochim Biophys Acta 1376:401–416

    Article  CAS  Google Scholar 

  17. Dumas F, Lebrun MC, Tocanne J-F (1999) Is the protein/lipid hydrophobic matching principle relevant to membrane organization and functions? FEBS Lett 458:271–277

    Article  CAS  Google Scholar 

  18. Andersen OS, Koeppe II RE (2007) Bilayer thickness and membrane protein function: an energetic perspective. Annu Rev Biophys Biomol Struct 36:107–130

    Article  CAS  Google Scholar 

  19. Rao BD, Shrivastava S, Chattopadhyay A (2017) Hydrophobic mismatch in membranes: when the tail matters. In: Chattopadhyay A (ed) Membrane organization and dynamics. Springer, Heidelberg, pp 375–387

    Chapter  Google Scholar 

  20. van der Wel PCA, Pott T, Morein S, Greathouse DV, Koeppe II RE, Killian JA (2000) Tryptophan-anchored transmembrane peptides promote formation of nonlamellar phases in phosphatidylethanolamine model membranes in a mismatch-dependent manner. Biochemistry 39:3124–3133

    Article  CAS  Google Scholar 

  21. Morein S, Koeppe II RE, Lindblom G, de Kruijff B, Killian JA (2000) The effect of peptide/lipid hydrophobic mismatch on the phase behavior of model membranes mimicking the lipid composition in Escherichia coli membranes. Biophys J 78:2475–2485

    Article  CAS  Google Scholar 

  22. de Planque MRR, Bonev BB, Demmers JAA, Greathouse DV, Koeppe II RE, Separovic F, Watts A, Killian JA (2003) Interfacial anchor properties of tryptophan residues in transmembrane peptides can dominate over hydrophobic matching effects in peptide-lipid interactions. Biochemistry 42:5341–5348

    Article  CAS  Google Scholar 

  23. de Planque MRR, Kruijtzer JAW, Liskamp RMJ, Marsh D, Greathouse DV, Koeppe II RE, de Kruijff B, Killian JA (1999) Different membrane anchoring positions of tryptophan and lysine in synthetic transmembrane α-helical peptides. J Biol Chem 274:20839–20846

  24. de Planque MRR, Boots J-WP, Rijkers DTS, Liskamp RMJ, Greathouse DV, Killian JA (2002) The effects of hydrophobic mismatch between phosphatidylcholine bilayers and transmembrane α-helical peptides depend on the nature of interfacially exposed aromatic and charged residues. Biochemistry 41:8396–8404

    Article  CAS  Google Scholar 

  25. Ӧzdirekcan S, Rijkers DTS, Liskamp RMJ, Killian JA (2005) Influence of flanking residues on tilt and rotation angles of transmembrane peptides in lipid bilayers. A solid-state 2H NMR study. Biochemistry 44:1004–1012

    Article  Google Scholar 

  26. Mortazavi A, Rajagopalan V, Sparks KA, Greathouse DV, Koeppe II RE (2016) Juxta-terminal helix unwinding as a stabilizing factor to modulate the dynamics of transmembrane helices. ChemBioChem 17:462–465

    Article  CAS  Google Scholar 

  27. Ghosh AK, Rukmini R, Chattopadhyay A (1997) Modulation of tryptophan environment in membrane-bound melittin by negatively charged phospholipids: implications in membrane organization and function. Biochemistry 36:14291–14305

    Article  CAS  Google Scholar 

  28. Rawat SS, Kelkar DA, Chattopadhyay A (2004) Monitoring gramicidin conformations in membranes: a fluorescence approach. Biophys J 87:831–843

    Article  CAS  Google Scholar 

  29. Chattopadhyay A, Rawat SS, Greathouse DV, Kelkar DA, Koeppe II RE (2008) Role of tryptophan residues in gramicidin channel organization and function. Biophys J 95:166–175

    Article  CAS  Google Scholar 

  30. Chaudhuri A, Haldar S, Sun H, Koeppe RE II, Chattopadhyay A (2014) Importance of indole N-H hydrogen bonding in the organization and dynamics of gramicidin channels. Biochim Biophys Acta 1838:419–428

    Article  CAS  Google Scholar 

  31. Mukherjee S, Chattopadhyay A (1995) Wavelength-selective fluorescence as a novel tool to study organization and dynamics in complex biological systems. J Fluoresc 5:237–246

    Article  CAS  Google Scholar 

  32. Demchenko AP (2008) Site-selective red-edge effects. Methods Enzymol 450:59–78

    Article  CAS  Google Scholar 

  33. Haldar S, Chaudhuri A, Chattopadhyay A (2011) Organization and dynamics of membrane probes and proteins utilizing the red edge excitation shift. J Phys Chem B 115:5693–5706

    Article  CAS  Google Scholar 

  34. Chattopadhyay A, Haldar S (2014) Dynamic insight into protein structure utilizing red edge excitation shift. Acc Chem Res 47:12–19

    Article  CAS  Google Scholar 

  35. Gleason NJ, Vostrikov VV, Greathouse DV, Grant CV, Opella SJ, Koeppe II RE (2012) Tyrosine replacing tryptophan as an anchor in GWALP peptides. Biochemistry 51:2044–2053

    Article  CAS  Google Scholar 

  36. Baron CB, Coburn RF (1984) Comparison of two copper reagents for detection of saturated and unsaturated neutral lipids by charring densitometry. J Liq Chromatogr 7:2793–2801

    Article  CAS  Google Scholar 

  37. McClare CWF (1971) An accurate and convenient organic phosphorus assay. Anal Biochem 39:527–530

    Article  CAS  Google Scholar 

  38. Chen GC, Yang JT (1977) Two-point calibration of circular dichrometer with d-10-camphorsulfonic acid. Anal Lett 10:1195–1207

    Article  CAS  Google Scholar 

  39. Chakraborty H, Chattopadhyay A (2017) Sensing tryptophan microenvironment of amyloid protein utilizing walvelength-selective fluorescence approach. J Fluoresc 27:1995–2000

    Article  CAS  Google Scholar 

  40. Rao BD, Chakraborty H, Keller S, Chattopadhyay A (2018) Aggregation behavior of pHLIP in aqueous solution at low concentrations: a fluorescence study. J Fluoresc 28:967–973

    Article  CAS  Google Scholar 

  41. Lakowicz JR (2006) Principles of fluorescence spectroscopy. 3rd edn. Springer, New York

    Book  Google Scholar 

  42. Kučerka N, Nieh M-P, Katsaras J (2011) Fluid phase lipid areas and bilayer thicknesses of commonly used phosphatidylcholines as a function of temperature. Biochim Biophys Acta 1808:2761–2771

    Article  Google Scholar 

  43. de Planque MRR, Goormaghtigh E, Greathouse DV, Koeppe II RE, Kruijtzer JAW, Liskamp RMJ, de Kruijff B, Killian JA (2001) Sensitivity of single membrane-spanning α-helical peptides to hydrophobic mismatch with a lipid bilayer: effects on backbone structure, orientation, and extent of membrane incorporation. Biochemistry 40:5000–5010

    Article  CAS  Google Scholar 

  44. Mukherjee S, Chattopadhyay A (1994) Motionally restricted tryptophan environments at the peptide-lipid interface of gramicidin channels. Biochemistry 33:5089–5097

    Article  CAS  Google Scholar 

  45. Prendergast FG (1991) Time-resolved fluorescence techniques: methods and applications in biology. Curr Opin Struct Biol 1:1054–1059

    Article  CAS  Google Scholar 

  46. Pal S, Aute R, Sarkar P, Bose S, Deshmukh MV, Chattopadhyay A (2018) Constrained dynamics of the sole tryptophan in the third intracellular loop of the serotonin1A receptor. Biophys Chem 240:34–41

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by intramural research grants from the Council of Scientific and Industrial Research, Govt. of India. A.C. gratefully acknowledges support from SERB Distinguished Fellowship (Department of Science and Technology, Govt. of India). R.E.K. acknowledges support from MCB grant 1713242 from the United States National Science Foundation. S.P. thanks the University Grants Commission for the award of a Senior Research Fellowship. We thank members of the Chattopadhyay laboratory for their comments and discussions.

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Authors and Affiliations

  1. CSIR-Indian Institute of Chemical Technology, Uppal Road, Hyderabad, 500 007, India

    Sreetama Pal

  2. Academy of Scientific and Innovative Research, Ghaziabad, India

    Sreetama Pal & Amitabha Chattopadhyay

  3. Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, 72701, USA

    Roger E. Koeppe II

  4. CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007, India

    Amitabha Chattopadhyay

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  1. Sreetama Pal
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  2. Roger E. Koeppe II
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Correspondence to Amitabha Chattopadhyay.

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Pal, S., Koeppe, R.E. & Chattopadhyay, A. Wavelength-Selective Fluorescence of a Model Transmembrane Peptide: Constrained Dynamics of Interfacial Tryptophan Anchors. J Fluoresc 28, 1317–1323 (2018). https://doi.org/10.1007/s10895-018-2293-5

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