Abstract
The interaction between seminalplasmin, an antimicrobial protein from bull semen, and lipid bilayers has been investigated. The fluorescence of the single tryptophan residue of the protein was measured. In the presence of phosphatidylcholine or phosphatidic acid bilayer vesicles the fluoresence maximum was shifted to shorter wavelengths, indicating transfer of the tryptophan to a more apolar environment. Circular dichroism spectra show an increased α-helical content for the protein in the presence of lipid. Quenching experiments clearly show the incorporation of the protein with the tryptophan localized near the bilayer surface. The shift of the tryptophan fluorescence emission was used to monitor the lipid phase transition in phosphatidylcholine membranes.
Similar content being viewed by others
Abbreviations
- TEMPOL:
-
2,2,6,6-Tetramethyl-4-hydroxy-piperidine-1-oxyl
- DMPC:
-
1,2-Dimyristoylphosphatidylcholine
- DMPA:
-
1,2-Dimyristoylphosphatidic acid
- SL 5:
-
2-(3-Carboxypropyl)-4,4-dimethyl-2-tridecyl-3-oxazolidinoxyl
- SL 12:
-
2-(10-Carboxydecyl)-4,4-dimethyl-2-hexyl-3-oxazolinoxyl
References
Dasseux JL, Faucon JF, Lafleur M, Pezolet M, Dufourcq J (1984) A restatement of melittin-induced effects on the thermotropism of zwitterionic phospholipids. Biochim Biophys Acta 775:37–50
Dufourq J, Faucon JF, Lussan C, Bernon R (1975) Study of lipid-protein interaction in membrane models: Intrinsic fluorescence of cytochrome b 5-phospholipid complexes. FEBS Lett 57:112–116
Galla HJ, Sackmann E, (1975a) Chemically induced phase separation in mixed vesicles containing phosphatidic acid. An optical study. J Am Chem Soc 97:4114–4120
Galla HJ, Sackmann E (1975b) Chemically induced lipid phase separation in model membranes containing charged lipids: A spin label study. Biochim Biophys Acta 401:509–529
Gietzen K, Bader H (1985) Effects of calmodulin antagonists on Ca2+-transport ATPase. In: Hidaka H, Hartshorne DJ (eds) Calmodulin antagonists and cellular physiology. Academic Press, New York, pp 347–362
Hartmann W, Galla HJ (1978) Binding of poly-lysine to charged bilayer membranes. Molecular organization of a lipid-peptide complex. Biochim Biophys Acta 509:474–490
Kapitza HG, Rüppel D, Galla HJ, Sackmann E (1984) Lateral diffusion of lipids and proteins in solid phosphatidylcholine bilayers: the role of structural defects. Biophys J 45:577–587
Luisetti J, Möhwald H, Galla HJ (1979) Monitoring the location profile of fluorophores in phosphatidylcholine bilayers by the use of paramagnetic quenching. Biochim Biophys Acta 552:519–530
Marsh D, Wattes A, Knowles PF (1976) Evidence for phase boundary lipid—Permeability of tempocholine into DML-vesicles at the phase transition temperature. Biochemistry 15:3570–3578
Mollay CH, Kreil G, Berger H (1976) Action of phospholipases on the cytoplasmic membrane of Escherichia coli. Stimulation by melittin. Biochim Biophys Acta 426:317–324
Mushayakarara I, Levin IW (1984) Effects of polypeptidephospholipid interactions on bilayer reorganizations. Raman spectroscopic study of the binding of polymyxin B to dimyristoylphosphatidic acid and dimyristoylphosphatidylcholine dispersions. Biochim Biophys Acta 769:585–595
Posch M, Rakusch U, Mollay Ch, Laggner P (1983) Cooperative effects in the interaction between melittin and phosphatidylcholine model membranes. J Biol Chem 258:1761–1766
Reddy ESP, Bharga PM (1979) Seminalplasmin — an antimicrobial protein from bovine seminalplasma which acts in E. coli by specific inhibition of rRNA synthesis. Nature 279:725–728
Rüppel D, Kapitza HG, Galla HJ, Sixl F, Sackmann E (1982) On the microstructure and the phasediagram of dimyristoylphosphatidylcholine-glycophorin bilayers. Biochim Biophys Acta 692:1–17
Scheit KH, Zimmer M (1984) Seminalplasma, an antimicrobial protein from bull semen, inhibits gene expression in E. coli. Biochim Biophys Acta 781:187–191
Scheit KH, Reddy ESP, Bhargara PM (1979) Seminalplasma is a potent inhibitor of E. coli RNA polymerase in vitro. Nature 279:728–732
Sixl F, Galla HJ (1979). Cooperative lipid-protein-interaction: Effect of pH and ionic strength on polymyxin to phosphatidic acid membranes. Biochim Biophys Acta 557:320–330
Sixl F, Galla HJ (1981) Polymyxin interaction with negatively charged lipid bilayer membranes and the competition effect of Ca2+-ions. Biochim Biophys Acta 643:626–635
Sixl F, Galla HJ (1982) Calorimetric investigation of polymyxin-binding to phosphatidic acid bilayers. Biochim Biophys Acta 693:466–478
Szoka F, Papahadjopoulos D (1978) Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse phase evaporation. Proc Natl Acad Sci USA 75:4194–4198
Theil R, Scheit KH (1983a) Amino acid sequence of seminalplasmin, an antimicrobial protein from bull semen. EMBO J 2:1159–1163
Theil R, Scheit KH (1983b) Purification and characterization of seminalplasmin, an antimicrobial protein from bull semen. Hoppe-Seyler's Z Physiol Chem 364:1003–1009
Träuble H, Eibl HJ (1974) Electrostatic effects on lipid phase transitions: Membrane structure and ionic environment. Proc Natl Acad Sci USA 71:214–219
Venkow P, Scheit KH (1984) Effects of seminalplasmin on rRNA synthesis in Saccharomyces cerevisiae. FEBS Lett 172:21–24
Vogel H (1981) Incorporation of melittin into phosphatidylcholine bilayers. FEBS Lett 134:37–42
Vogel H, Jähnig F, Hoffmann V, Stümpel J (1983) The orientation of melittin in lipid membranes. A polarized infrared spectroscopy study. Biochim Biophys Acta 733:201–209
Williams JC, Bell RM (1972) Membrane matrix disruption by melittin. Biochim Biophys Acta 288:255–262
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Galla, HJ., Warncke, M. & Scheit, KH. Incorporation of the antimicrobial protein seminalplasmin into lipid bilayer membranes. Eur Biophys J 12, 211–216 (1985). https://doi.org/10.1007/BF00253847
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00253847