Skip to main content
SpringerLink
Log in

Kinetics of proton transfer in a green fluorescent protein: A laser-induced pH jump study

  • Published:
Journal of Chemical Sciences Aims and scope Submit manuscript

Abstract

The sub-millisecond protonation dynamics of the chromophore in S65T mutant form of the green fluorescent protein (GFP) was tracked after a rapid pH jump following laser-induced proton release from the caged photolabile compoundo-nitrobenzaldehyde. Following a jump in pH from 8 to 5 (which is achieved within 2 μs), the fluorescence of S65T GFP decreased as a single exponential with a time constant of ∼90 μs. This decay is interpreted as the conversion of the deprotonated fluorescent GFP chromophore to a protonated non-fluorescent species. The protonation kinetics showed dependence on the bulk viscosity of the solvent, and therefore implicates bulk solvent-controlled protein dynamics in the protonation process. The protonation is proposed to be a sequential process involving two steps: (a) proton transfer from solvent to the chromophore, and (b) internal structural rearrangements to stabilize a protonated chromophore. The possible implications of these observations to protein dynamics in general is discussed

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Fenimore P W, Frauenfelder H, McMahon B H and Parak F G 2002Proc. Natl. Acad. Sci. (USA) 99 16047

    Article  CAS  Google Scholar 

  2. Kay L E 1998Nature Struct. Biol. (NMR Suppl.) 5 513

    Article  CAS  Google Scholar 

  3. Stock A 1999Nature (London) 400 221

    Article  CAS  Google Scholar 

  4. Feher V A and Cavanagh J 1999Nature (London) 400 289

    Article  CAS  Google Scholar 

  5. Nicholson L K, Yamazaki T, Torchia D A, Grzesiek S, Bax A, Stahl S J, Kaufman J D, Wingfield P T, Lam P Y and Jadhav P K 1995Nature Struct. Biol. 2 274

    Article  CAS  Google Scholar 

  6. Lee A L, Kinnear S A and Wand A J 2000Nature Struct. Biol. 7 72

    Article  CAS  Google Scholar 

  7. Luecke H, Schobert B, Richter H T, Cartailler J P and Lanyi J K 1999Science 286 255

    Article  CAS  Google Scholar 

  8. Subramaniam S and Henderson 2000Nature (London) 406 653

    Article  CAS  Google Scholar 

  9. Abrahams J P, Leslie A G, Lutte R and Walker J E 1994Nature (London) 370 621

    Article  CAS  Google Scholar 

  10. Iwata S, Ostermier C, Ludwig B and Michel H 1995Nature (London) 376 660

    Article  CAS  Google Scholar 

  11. Tsukihara T, Aoyama H, Yamashita E, Tomizaki T, Yamaguchi H, Shinzawa-Itoh, Nakashima R, Yaono R and Yoshikawa S 1996Science 272 1136

    Article  Google Scholar 

  12. Cavanagh J and Akke M 2000Nature Struct. Biol. 7 11

    Article  CAS  Google Scholar 

  13. Wand J A 2001Nature Struct. Biol. 8 926

    Article  CAS  Google Scholar 

  14. Shimomura O, Johnson F H and Saiga Y 1962J. Cell. Comp. Physiol. 59 223

    Article  CAS  Google Scholar 

  15. Tsien R Y 1998Annu. Rev. Biochem. 67 509

    Article  CAS  Google Scholar 

  16. Ormo M, Cubitt A B, Kallio K, Gross L A, Tsien R Y and Remington S J 1996Science 273 1392

    Article  Google Scholar 

  17. Yang F, Moss L G and Phillips G N Jr 1996Nature Biotechnol. 14 1246

    Article  Google Scholar 

  18. Heim R, Prasher D C and Tsien R Y 1994Proc. Natl. Acad. Sci. (USA) 91 12501

    Article  CAS  Google Scholar 

  19. Brejc K, Sixma T K, Kitts P A, Kain S R, Tsien R Y, Ormo M and Remington S J 1997Proc. Natl. Acad. Sci. (USA) 94 2306

    Article  CAS  Google Scholar 

  20. Elsliger M, Wachter R M, Hanson G T, Kallio K and Remington S J 1999Biochemistry 38 5296

    Article  CAS  Google Scholar 

  21. Haupts U, Maiti S, Schwille P and Webb WW 1998Proc. Natl. Acad. Sci. (USA) 95 13573

    Article  CAS  Google Scholar 

  22. Kneen M, Farinas J, Li Y and Verkman A S 1998Biophys. J. 74 1591

    Article  CAS  Google Scholar 

  23. Viappiani C, Bonettu G, Carcelli M, Ferrari F and Sternieri A 1998Rev. Sci. Instrum. 69 270

    Article  CAS  Google Scholar 

  24. Abbruzzetti S, Crema E, Masino L, Veeli A, Viappiani C, Small J R, Libertini L J and Small E W 2000Biophys. J. 78 405

    CAS  Google Scholar 

  25. Barth A and Corriedaggo JET 2002Biophys. J. 83 2864

    CAS  Google Scholar 

  26. Chattoraj M, King B A, Bublitz G U and Boxer S G 1996Proc. Natl. Acad. Sci. (USA) 93 8362

    Article  Google Scholar 

  27. Swaminathan R, Hoang C P and Verkman A S 1997Biophys. J. 72 1900

    CAS  Google Scholar 

  28. Gutman M and Nachliel E 1990Biochim. Biophys. Acta 1015 391

    Article  CAS  Google Scholar 

  29. George M V and Scaiano J C 1980J. Phys. Chem. 84 492

    Article  CAS  Google Scholar 

  30. Chen K, Hirst J, Camba R, Bonagura C A, Stout C D, Burgess B K and Armstrong F A 2000Nature (London) 405 814

    Article  CAS  Google Scholar 

  31. Lill M A and Helms V 2002Proc. Natl. Acad. Sci. (USA) 99 2778

    Article  CAS  Google Scholar 

  32. Pomes R and Roux B 1996J. Phys. Chem. 100 2519

    Article  Google Scholar 

  33. Brewer M L, Schmitt U W and Voth G A 2001Biophys. J. 80 1691

    CAS  Google Scholar 

  34. Gavish B and Yedgar S 1995Protein-solvent interactions (ed.) R B Gregory (New York: Marcel Dekker) p. 343

    Google Scholar 

  35. Gregory R G, Rosenberg A, Knox D and Percy A J 1990Biopolymers 29 1175

    Article  CAS  Google Scholar 

  36. Ansari A, Lones C M, Henry E R, Hofrichter J and Eaton W A 1992Science 256 1796

    Article  CAS  Google Scholar 

  37. Kramers H A 1940Physica (Utrecht) 7 284

    Article  CAS  Google Scholar 

  38. Hanggi P, Talkner P and Borkovec M 1990Rev. Mod. Phys. 62 251

    Article  Google Scholar 

Download references

Author information

Author notes

  1. Roop Mallik

    Present address: Department of Developmental & Cell Biology, University of California Irvine, 2302 Natural Sciences 1, 92697, Irvine, CA, USA

Authors and Affiliations

  1. National Centre for Biological Sciences, Tata Institute of Fundamental Research, GKVK Campus, 560 065, Bangalore, India

    Roop Mallik & Jayant B. Udgaonkar

  2. Department of Chemical Sciences, Tata Institute of Fundamental Research, 400 005, Mumbai, India

    G. Krishnamoorthy

Authors
  1. Roop Mallik
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jayant B. Udgaonkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Krishnamoorthy
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. Krishnamoorthy.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mallik, R., Udgaonkar, J.B. & Krishnamoorthy, G. Kinetics of proton transfer in a green fluorescent protein: A laser-induced pH jump study. J Chem Sci 115, 307–317 (2003). https://doi.org/10.1007/BF02704222

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02704222

Keywords

Search

Navigation