Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jun 11;93(12):5872–5876. doi: 10.1073/pnas.93.12.5872

Conformational trapping in a membrane environment: a regulatory mechanism for protein activity?

S Arumugam 1, S Pascal 1, C L North 1, W Hu 1, K C Lee 1, M Cotten 1, R R Ketchem 1, F Xu 1, M Brenneman 1, F Kovacs 1, F Tian 1, A Wang 1, S Huo 1, T A Cross 1
PMCID: PMC39154  PMID: 8650185

Abstract

Functional regulation of proteins is central to living organisms. Here it is shown that a nonfunctional conformational state of a polypeptide can be kinetically trapped in a lipid bilayer environment. This state is a metastable structure that is stable for weeks just above the phase transition temperature of the lipid. When the samples are incubated for several days at 68 degrees C, 50% of the trapped conformation converts to the minimum-energy functional state. This result suggests the possibility that another mechanism for functional regulation of protein activity may be available for membrane proteins: that cells may insert proteins into membranes in inactive states pending the biological demand for protein function.

Full text

PDF
5872

Images in this article

5873Fig. 1
on p.5873
5875Fig. 4
on p.5875

Selected References

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

  1. Affleck R., Xu Z. F., Suzawa V., Focht K., Clark D. S., Dordick J. S. Enzymatic catalysis and dynamics in low-water environments. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):1100–1104. doi: 10.1073/pnas.89.3.1100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker D., Agard D. A. Kinetics versus thermodynamics in protein folding. Biochemistry. 1994 Jun 21;33(24):7505–7509. doi: 10.1021/bi00190a002. [DOI] [PubMed] [Google Scholar]
  3. Bañ M. C., Braco L., Abad C. HPLC study on the 'history' dependence of gramicidin A conformation in phospholipid model membranes. FEBS Lett. 1989 Jun 19;250(1):67–71. doi: 10.1016/0014-5793(89)80686-6. [DOI] [PubMed] [Google Scholar]
  4. Douliez J. P., Léonard A., Dufourc E. J. Restatement of order parameters in biomembranes: calculation of C-C bond order parameters from C-D quadrupolar splittings. Biophys J. 1995 May;68(5):1727–1739. doi: 10.1016/S0006-3495(95)80350-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fields C. G., Fields G. B., Noble R. L., Cross T. A. Solid phase peptide synthesis of 15N-gramicidins A, B, and C and high performance liquid chromatographic purification. Int J Pept Protein Res. 1989 Apr;33(4):298–303. doi: 10.1111/j.1399-3011.1989.tb01285.x. [DOI] [PubMed] [Google Scholar]
  6. Ketchem R. R., Hu W., Cross T. A. High-resolution conformation of gramicidin A in a lipid bilayer by solid-state NMR. Science. 1993 Sep 10;261(5127):1457–1460. doi: 10.1126/science.7690158. [DOI] [PubMed] [Google Scholar]
  7. Killian J. A., Prasad K. U., Hains D., Urry D. W. The membrane as an environment of minimal interconversion. A circular dichroism study on the solvent dependence of the conformational behavior of gramicidin in diacylphosphatidylcholine model membranes. Biochemistry. 1988 Jun 28;27(13):4848–4855. doi: 10.1021/bi00413a040. [DOI] [PubMed] [Google Scholar]
  8. Koeppe R. E., 2nd, Killian J. A., Greathouse D. V. Orientations of the tryptophan 9 and 11 side chains of the gramicidin channel based on deuterium nuclear magnetic resonance spectroscopy. Biophys J. 1994 Jan;66(1):14–24. doi: 10.1016/S0006-3495(94)80748-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Langs D. A. Three-dimensional structure at 0.86 A of the uncomplexed form of the transmembrane ion channel peptide gramicidin A. Science. 1988 Jul 8;241(4862):188–191. doi: 10.1126/science.2455345. [DOI] [PubMed] [Google Scholar]
  10. LoGrasso P. V., Moll F., 3rd, Cross T. A. Solvent history dependence of gramicidin A conformations in hydrated lipid bilayers. Biophys J. 1988 Aug;54(2):259–267. doi: 10.1016/S0006-3495(88)82955-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Malcolm B. A., Wilson K. P., Matthews B. W., Kirsch J. F., Wilson A. C. Ancestral lysozymes reconstructed, neutrality tested, and thermostability linked to hydrocarbon packing. Nature. 1990 May 3;345(6270):86–89. doi: 10.1038/345086a0. [DOI] [PubMed] [Google Scholar]
  12. Nicholson L. K., Cross T. A. Gramicidin cation channel: an experimental determination of the right-handed helix sense and verification of beta-type hydrogen bonding. Biochemistry. 1989 Nov 28;28(24):9379–9385. doi: 10.1021/bi00450a019. [DOI] [PubMed] [Google Scholar]
  13. O'Connell A. M., Koeppe R. E., 2nd, Andersen O. S. Kinetics of gramicidin channel formation in lipid bilayers: transmembrane monomer association. Science. 1990 Nov 30;250(4985):1256–1259. doi: 10.1126/science.1700867. [DOI] [PubMed] [Google Scholar]
  14. Pascal S. M., Cross T. A. High-resolution structure and dynamic implications for a double-helical gramicidin A conformer. J Biomol NMR. 1993 Sep;3(5):495–513. doi: 10.1007/BF00174606. [DOI] [PubMed] [Google Scholar]
  15. Pascal S. M., Cross T. A. Structure of an isolated gramicidin A double helical species by high-resolution nuclear magnetic resonance. J Mol Biol. 1992 Aug 20;226(4):1101–1109. doi: 10.1016/0022-2836(92)91055-t. [DOI] [PubMed] [Google Scholar]
  16. Popot J. L., Engelman D. M. Membrane protein folding and oligomerization: the two-stage model. Biochemistry. 1990 May 1;29(17):4031–4037. doi: 10.1021/bi00469a001. [DOI] [PubMed] [Google Scholar]
  17. Rodrigueza W. V., Wheeler J. J., Klimuk S. K., Kitson C. N., Hope M. J. Transbilayer movement and net flux of cholesterol and cholesterol sulfate between liposomal membranes. Biochemistry. 1995 May 9;34(18):6208–6217. doi: 10.1021/bi00018a025. [DOI] [PubMed] [Google Scholar]
  18. Shoichet B. K., Baase W. A., Kuroki R., Matthews B. W. A relationship between protein stability and protein function. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):452–456. doi: 10.1073/pnas.92.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shon K. J., Kim Y., Colnago L. A., Opella S. J. NMR studies of the structure and dynamics of membrane-bound bacteriophage Pf1 coat protein. Science. 1991 May 31;252(5010):1303–1305. doi: 10.1126/science.1925542. [DOI] [PubMed] [Google Scholar]
  20. Smith R., Separovic F., Milne T. J., Whittaker A., Bennett F. M., Cornell B. A., Makriyannis A. Structure and orientation of the pore-forming peptide, melittin, in lipid bilayers. J Mol Biol. 1994 Aug 19;241(3):456–466. doi: 10.1006/jmbi.1994.1520. [DOI] [PubMed] [Google Scholar]
  21. Ulrich A. S., Watts A., Wallat I., Heyn M. P. Distorted structure of the retinal chromophore in bacteriorhodopsin resolved by 2H-NMR. Biochemistry. 1994 May 10;33(18):5370–5375. doi: 10.1021/bi00184a003. [DOI] [PubMed] [Google Scholar]
  22. Urry D. W., Long M. M., Jacobs M., Harris R. D. Conformation and molecular mechanisms of carriers and channels. Ann N Y Acad Sci. 1975 Dec 30;264:203–220. doi: 10.1111/j.1749-6632.1975.tb31484.x. [DOI] [PubMed] [Google Scholar]
  23. Veatch W. R., Fossel E. T., Blout E. R. The conformation of gramicidin A. Biochemistry. 1974 Dec 17;13(26):5249–5256. doi: 10.1021/bi00723a001. [DOI] [PubMed] [Google Scholar]
  24. Wallace B. A. Gramicidin channels and pores. Annu Rev Biophys Biophys Chem. 1990;19:127–157. doi: 10.1146/annurev.bb.19.060190.001015. [DOI] [PubMed] [Google Scholar]
  25. Zaks A., Klibanov A. M. The effect of water on enzyme action in organic media. J Biol Chem. 1988 Jun 15;263(17):8017–8021. [PubMed] [Google Scholar]
  26. Zhang Z., Pascal S. M., Cross T. A. A conformational rearrangement in gramicidin A: from a double-stranded left-handed to a single-stranded right-handed helix. Biochemistry. 1992 Sep 22;31(37):8822–8828. doi: 10.1021/bi00152a019. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES