Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Jul 15;365(Pt 2):471–479. doi: 10.1042/BJ20011853

Aerobic sn-glycerol-3-phosphate dehydrogenase from Escherichia coli binds to the cytoplasmic membrane through an amphipathic alpha-helix.

Antje-Christine Walz 1, Rudy A Demel 1, Ben de Kruijff 1, Rupert Mutzel 1
PMCID: PMC1222694  PMID: 11955283

Abstract

sn-Glycerol-3-phosphate dehydrogenase (GlpD) from Escherichia coli is a peripheral membrane enzyme involved in respiratory electron transfer. For it to display its enzymic activity, binding to the inner membrane is required. The way the enzyme interacts with the membrane and how this controls activity has not been elucidated. In the present study we provide evidence for direct protein-lipid interaction. Using the monolayer technique, we observed insertion of GlpD into lipid monolayers with a clear preference for anionic phospholipids. GlpD variants with point mutations in their predicted amphipathic helices showed a decreased ability to penetrate anionic phospholipid monolayers. From these data we propose that membrane binding of GlpD occurs by insertion of an amphipathic helix into the acyl-chain region of lipids mediated by negatively charged phospholipids.

Full Text

The Full Text of this article is available as a PDF (234.3 KB).

Selected References

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

  1. Austin D., Larson T. J. Nucleotide sequence of the glpD gene encoding aerobic sn-glycerol 3-phosphate dehydrogenase of Escherichia coli K-12. J Bacteriol. 1991 Jan;173(1):101–107. doi: 10.1128/jb.173.1.101-107.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bernstein L. S., Grillo A. A., Loranger S. S., Linder M. E. RGS4 binds to membranes through an amphipathic alpha -helix. J Biol Chem. 2000 Jun 16;275(24):18520–18526. doi: 10.1074/jbc.M000618200. [DOI] [PubMed] [Google Scholar]
  4. Boos W. Binding protein-dependent ABC transport system for glycerol 3-phosphate of Escherichia coli. Methods Enzymol. 1998;292:40–51. doi: 10.1016/s0076-6879(98)92006-7. [DOI] [PubMed] [Google Scholar]
  5. Borgnia M. J., Agre P. Reconstitution and functional comparison of purified GlpF and AqpZ, the glycerol and water channels from Escherichia coli. Proc Natl Acad Sci U S A. 2001 Feb 20;98(5):2888–2893. doi: 10.1073/pnas.051628098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Breukink E., Demel R. A., de Korte-Kool G., de Kruijff B. SecA insertion into phospholipids is stimulated by negatively charged lipids and inhibited by ATP: a monolayer study. Biochemistry. 1992 Feb 4;31(4):1119–1124. doi: 10.1021/bi00119a021. [DOI] [PubMed] [Google Scholar]
  7. Demel R. A., Geurts van Kessel W. S., Zwaal R. F., Roelofsen B., van Deenen L. L. Relation between various phospholipase actions on human red cell membranes and the interfacial phospholipid pressure in monolayers. Biochim Biophys Acta. 1975 Sep 16;406(1):97–107. doi: 10.1016/0005-2736(75)90045-0. [DOI] [PubMed] [Google Scholar]
  8. Demel R. A. Monomolecular layers in the study of biomembranes. Subcell Biochem. 1994;23:83–120. doi: 10.1007/978-1-4615-1863-1_3. [DOI] [PubMed] [Google Scholar]
  9. Dowhan W. Molecular basis for membrane phospholipid diversity: why are there so many lipids? Annu Rev Biochem. 1997;66:199–232. doi: 10.1146/annurev.biochem.66.1.199. [DOI] [PubMed] [Google Scholar]
  10. Eisenberg D., Schwarz E., Komaromy M., Wall R. Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol. 1984 Oct 15;179(1):125–142. doi: 10.1016/0022-2836(84)90309-7. [DOI] [PubMed] [Google Scholar]
  11. Fishov I., Woldringh C. L. Visualization of membrane domains in Escherichia coli. Mol Microbiol. 1999 Jun;32(6):1166–1172. doi: 10.1046/j.1365-2958.1999.01425.x. [DOI] [PubMed] [Google Scholar]
  12. Flower A. M. SecG function and phospholipid metabolism in Escherichia coli. J Bacteriol. 2001 Mar;183(6):2006–2012. doi: 10.1128/JB.183.6.2006-2012.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frishman D., Argos P. Incorporation of non-local interactions in protein secondary structure prediction from the amino acid sequence. Protein Eng. 1996 Feb;9(2):133–142. doi: 10.1093/protein/9.2.133. [DOI] [PubMed] [Google Scholar]
  14. Garner J., Crooke E. Membrane regulation of the chromosomal replication activity of E. coli DnaA requires a discrete site on the protein. EMBO J. 1996 Jul 1;15(13):3477–3485. [PMC free article] [PubMed] [Google Scholar]
  15. Garner J., Durrer P., Kitchen J., Brunner J., Crooke E. Membrane-mediated release of nucleotide from an initiator of chromosomal replication, Escherichia coli DnaA, occurs with insertion of a distinct region of the protein into the lipid bilayer. J Biol Chem. 1998 Feb 27;273(9):5167–5173. doi: 10.1074/jbc.273.9.5167. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gibrat J. F., Garnier J., Robson B. Further developments of protein secondary structure prediction using information theory. New parameters and consideration of residue pairs. J Mol Biol. 1987 Dec 5;198(3):425–443. doi: 10.1016/0022-2836(87)90292-0. [DOI] [PubMed] [Google Scholar]
  18. Havell E. A., Spitalny G. L. Production and characterization of anti-murine interferon-gamma sera. J Interferon Res. 1983;3(2):191–198. doi: 10.1089/jir.1983.3.191. [DOI] [PubMed] [Google Scholar]
  19. Heller K. B., Lin E. C., Wilson T. H. Substrate specificity and transport properties of the glycerol facilitator of Escherichia coli. J Bacteriol. 1980 Oct;144(1):274–278. doi: 10.1128/jb.144.1.274-278.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
  21. Holtman C. K., Pawlyk A. C., Meadow N. D., Pettigrew D. W. Reverse genetics of Escherichia coli glycerol kinase allosteric regulation and glucose control of glycerol utilization in vivo. J Bacteriol. 2001 Jun;183(11):3336–3344. doi: 10.1128/JB.183.11.3336-3344.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kung H. F., Henning U. Limiting availability of binding sites for dehydrogenases on the cell membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Apr;69(4):925–929. doi: 10.1073/pnas.69.4.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Larson T. J., Ehrmann M., Boos W. Periplasmic glycerophosphodiester phosphodiesterase of Escherichia coli, a new enzyme of the glp regulon. J Biol Chem. 1983 May 10;258(9):5428–5432. [PubMed] [Google Scholar]
  24. Larson T. J., Schumacher G., Boos W. Identification of the glpT-encoded sn-glycerol-3-phosphate permease of Escherichia coli, an oligomeric integral membrane protein. J Bacteriol. 1982 Dec;152(3):1008–1021. doi: 10.1128/jb.152.3.1008-1021.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Larson T. J., Ye S. Z., Weissenborn D. L., Hoffmann H. J., Schweizer H. Purification and characterization of the repressor for the sn-glycerol 3-phosphate regulon of Escherichia coli K12. J Biol Chem. 1987 Nov 25;262(33):15869–15874. [PubMed] [Google Scholar]
  26. Makise M., Mima S., Tsuchiya T., Mizushima T. Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids. J Biol Chem. 2000 Feb 11;275(6):4513–4518. doi: 10.1074/jbc.275.6.4513. [DOI] [PubMed] [Google Scholar]
  27. Mileykovskaya E., Dowhan W. Visualization of phospholipid domains in Escherichia coli by using the cardiolipin-specific fluorescent dye 10-N-nonyl acridine orange. J Bacteriol. 2000 Feb;182(4):1172–1175. doi: 10.1128/jb.182.4.1172-1175.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miller A., Wang L., Kendall D. A. Synthetic signal peptides specifically recognize SecA and stimulate ATPase activity in the absence of preprotein. J Biol Chem. 1998 May 8;273(19):11409–11412. doi: 10.1074/jbc.273.19.11409. [DOI] [PubMed] [Google Scholar]
  29. Norris V., Fishov I. Hypothesis: membrane domains and hyperstructures control bacterial division. Biochimie. 2001 Jan;83(1):91–97. doi: 10.1016/s0300-9084(00)01203-7. [DOI] [PubMed] [Google Scholar]
  30. Picot D., Loll P. J., Garavito R. M. The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1. Nature. 1994 Jan 20;367(6460):243–249. doi: 10.1038/367243a0. [DOI] [PubMed] [Google Scholar]
  31. Rietveld A. G., Koorengevel M. C., de Kruijff B. Non-bilayer lipids are required for efficient protein transport across the plasma membrane of Escherichia coli. EMBO J. 1995 Nov 15;14(22):5506–5513. doi: 10.1002/j.1460-2075.1995.tb00237.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Robinson J. J., Weiner J. H. The effect of amphipaths on the flavin-linked aerobic glycerol-3-phosphate dehydrogenase from Escherichia coli. Can J Biochem. 1980 Oct;58(10):1172–1178. doi: 10.1139/o80-157. [DOI] [PubMed] [Google Scholar]
  33. Rost B., Sander C. Prediction of protein secondary structure at better than 70% accuracy. J Mol Biol. 1993 Jul 20;232(2):584–599. doi: 10.1006/jmbi.1993.1413. [DOI] [PubMed] [Google Scholar]
  34. Schryvers A., Lohmeier E., Weiner J. H. Chemical and functional properties of the native and reconstituted forms of the membrane-bound, aerobic glycerol-3-phosphate dehydrogenase of Escherichia coli. J Biol Chem. 1978 Feb 10;253(3):783–788. [PubMed] [Google Scholar]
  35. Segrest J. P., Jones M. K., De Loof H., Brouillette C. G., Venkatachalapathi Y. V., Anantharamaiah G. M. The amphipathic helix in the exchangeable apolipoproteins: a review of secondary structure and function. J Lipid Res. 1992 Feb;33(2):141–166. [PubMed] [Google Scholar]
  36. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  37. Wang G., Peterkofsky A., Clore G. M. A novel membrane anchor function for the N-terminal amphipathic sequence of the signal-transducing protein IIAGlucose of the Escherichia coli phosphotransferase system. J Biol Chem. 2000 Dec 22;275(51):39811–39814. doi: 10.1074/jbc.C000709200. [DOI] [PubMed] [Google Scholar]
  38. Weiner J. H., Heppel L. A. Purification of the membrane-bound and pyridine nucleotide-independent L-glycerol 3-phosphate dehydrogenase from Escherichia coli. Biochem Biophys Res Commun. 1972 Jun 28;47(6):1360–1365. doi: 10.1016/0006-291x(72)90222-7. [DOI] [PubMed] [Google Scholar]
  39. Weiner J. H. The localization of glycerol-3-phosphate dehydrogenase in Escherichia coli. J Membr Biol. 1974;15(1):1–14. doi: 10.1007/BF01870078. [DOI] [PubMed] [Google Scholar]
  40. Yang B., Larson T. J. Action at a distance for negative control of transcription of the glpD gene encoding sn-glycerol 3-phosphate dehydrogenase of Escherichia coli K-12. J Bacteriol. 1996 Dec;178(24):7090–7098. doi: 10.1128/jb.178.24.7090-7098.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zwaig N., Kistler W. S., Lin E. C. Glycerol kinase, the pacemaker for the dissimilation of glycerol in Escherichia coli. J Bacteriol. 1970 Jun;102(3):753–759. doi: 10.1128/jb.102.3.753-759.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. de Leeuw E., te Kaat K., Moser C., Menestrina G., Demel R., de Kruijff B., Oudega B., Luirink J., Sinning I. Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity. EMBO J. 2000 Feb 15;19(4):531–541. doi: 10.1093/emboj/19.4.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. van Kraaij C., Breukink E., Noordermeer M. A., Demel R. A., Siezen R. J., Kuipers O. P., de Kruijff B. Pore formation by nisin involves translocation of its C-terminal part across the membrane. Biochemistry. 1998 Nov 17;37(46):16033–16040. doi: 10.1021/bi980931b. [DOI] [PubMed] [Google Scholar]
  44. van den Brink-van der Laan E., Dalbey R. E., Demel R. A., Killian J. A., de Kruijff B. Effect of nonbilayer lipids on membrane binding and insertion of the catalytic domain of leader peptidase. Biochemistry. 2001 Aug 14;40(32):9677–9684. doi: 10.1021/bi002903a. [DOI] [PubMed] [Google Scholar]

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

RESOURCES