Secondary structure of M13 coat protein in phospholipids studied by circular dichroism, Raman, and Fourier transform infrared spectroscopy.

There is considerable uncertainty about the precise secondary structure adopted by the M13 coat protein when embedded in a phospholipid bilayer. Circular dichroism (CD) spectroscopy suggests that a major change in the structure of the coat protein occurs upon membrane insertion. It is reported that the structure of the protein in the membrane has only about 50% alpha-helix, the rest being mainly in a beta-sheet conformation, whereas the protein is almost completely alpha-helical when intact in the phage. In this study we have undertaken a spectroscopic analysis using Fourier transform infrared, Raman, and CD spectroscopy to characterize the secondary structure of M13 coat protein when present in membranes consisting of dioleoylphosphatidylglycerol and dimyristoylphosphatidylglycerol. In sharp contrast to earlier CD studies, our results indicate that the coat protein in its membrane-embedded state has a very high alpha-helical content with virtually no beta-sheet structures present. This result indicates that the structures of the coat protein when intact in the phage or when embedded in the membrane are similar. Although our results differ from earlier CD studies, they are consistent with a recent NMR study, which showed that the M13 coat protein in sodium dodecyl sulfate micelles is primarily alpha-helical with no evidence for beta-sheet structure [Henry, G. D., & Sykes, B.D. (1992) Biochemistry 31, 5284-5297]. These results lead to the conclusion that the M13 coat protein can insert from the membrane-bound state into a virus particle with a similar secondary structure, without large energy implications.(ABSTRACT TRUNCATED AT 250 WORDS)

[1]  A. Visser,et al.  Analysis of time-resolved fluorescence anisotropy in lipid-protein systems , 2004, European Biophysics Journal.

[2]  H. Hoffmann-Berling,et al.  A coat protein from bacteriophage fd. I. Hydrodynamic measurements and biological characterization. , 1966, Journal of molecular biology.

[3]  D. Gray,et al.  A CD determination of the α‐helix contents of the coat proteins of four filamentous bacteriophages: fd, IKe, Pf1, and Pf3 , 1989, Biopolymers.

[4]  K. Valentine,et al.  Dynamics of fd coat protein in lipid bilayers. , 1987, Biochemistry.

[5]  L. Makowski,et al.  Three-dimensional structure of a cloning vector. X-ray diffraction studies of filamentous bacteriophage M13 at 7 A resolution. , 1992, Journal of molecular biology.

[6]  A model for fd phage penetration and assembly , 1991, FEBS letters.

[7]  H. Smilowitz.  Bacteriophage f1 Infection: Fate of the Parental Major Coat Protein , 1974, Journal of virology.

[8]  C. Tanford,et al.  Conformational states of a hydrophobic protein. The coat protein of fd bacteriophage. , 1978, Biochemistry.

[9]  R. Chambert,et al.  A Raman spectroscopic study on the interaction of an ion-channel protein with a phospholipid in a model membrane system (gramicidin A/L-alpha-lysophosphatidylcholine). , 1986, European journal of biochemistry.

[10]  A. Visser,et al.  Analysis of time-resolved fluorescence anisotropy in lipid-protein systems , 1990, European Biophysics Journal.

[11]  Structure and dynamics of detergent-solubilized M13 coat protein (an integral membrane protein) determined by 13C and 15N nuclear magnetic resonance spectroscopy. , 1990, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[12]  P. Haris,et al.  Conformational transitions in poly (L-lysine): studies using Fourier transform infrared spectroscopy , 1989 .

[13]  B. Sykes,et al.  Side-chain dynamics of a detergent-solubilized membrane protein: measurement of tryptophan and glutamine hydrogen-exchange rates in M13 coat protein by 1H NMR spectroscopy. , 1989, Biochemistry.

[14]  T. Poile,et al.  A NMR investigation on the interactions of the alpha-oligomeric form of the M13 coat protein with lipids, which mimic the Escherichia coli inner membrane. , 1991, Biochimica et biophysica acta.

[15]  Alain J. P. Alix,et al.  Determination of the secondary structure of proteins from the Raman amide I band: the reference intensity profiles method , 1987 .

[16]  R. Spruijt,et al.  Aggregation-related conformational change of the membrane-associated coat protein of bacteriophage M13. , 1989, Biochemistry.

[17]  R. Spruijt,et al.  The in situ aggregational and conformational state of the major coat protein of bacteriophage M13 in phospholipid bilayers mimicking the inner membrane of host Escherichia coli. , 1991, Biochemistry.

[18]  A. Ge,et al.  Tryptophan contributions to the unusual circular dichroism of fd bacteriophage. , 1992 .

[19]  S. Opella,et al.  Comparison of the dynamics of the membrane‐bound form of fd coat protein in micelles and in bilayers by solution and solid‐state nitrogen‐15 nuclear magnetic resonance spectroscopy , 1988, Proteins.

[20]  S. Opella,et al.  NMR studies of the structure and dynamics of membrane-bound bacteriophage Pf1 coat protein. , 1991, Science.

[21]  H. Mantsch,et al.  New insight into protein secondary structure from resolution-enhanced infrared spectra. , 1988, Biochimica et biophysica acta.

[22]  A. Alfsen,et al.  Comparative study of myelin proteolipid apoprotein solvation by multilayer membranes of synthetic DPPC and biological lipid extract from bovine brain. An FT-IR investigation. , 1989, Biochimie.

[23]  D. Marvin,et al.  Model-building studies of Inovirus: genetic variations on a geometric theme. , 1990, International journal of biological macromolecules.

[24]  R. Williams,et al.  Estimation of protein secondary structure from the laser Raman amide I spectrum. , 1983, Journal of molecular biology.

[25]  P. Wolber,et al.  Bilayer acyl chain dynamics and lipid-protein interaction: the effect of the M13 bacteriophage coat protein on the decay of the fluorescence anisotropy of parinaric acid. , 1982, Biophysical journal.

[26]  S. Provencher,et al.  Estimation of globular protein secondary structure from circular dichroism. , 1981, Biochemistry.

[27]  R. E. Webster,et al.  Evidence for a major conformational change of coat protein in assembly of fl bacteriophage , 1976, Nature.

[28]  B. Sykes,et al.  Fluorotyrosine M13 coat protein: fluorine-19 nuclear magnetic resonance study of the motional properties of an integral membrane protein in phospholipid vesicles. , 1978, Biochemistry.

[29]  B. Sykes,et al.  Hydrogen exchange kinetics in a membrane protein determined by 15N NMR spectroscopy: use of the INEPT experiment to follow individual amides in detergent-solubilized M13 coat protein. , 1990, Biochemistry.

[30]  R. E. Webster,et al.  Association of the major coat protein of fd bacteriophage with phospholipid vesicles. , 1978, Biochimica et biophysica acta.

[31]  G. Feigenson,et al.  Reconstitution of M13 bacteriophage coat protein. A new strategy to analyze configuration of the protein in the membrane. , 1985, Biochimica et biophysica acta.

[32]  P. Wolber,et al.  Protein-lipid interactions. Studies of the M13 coat protein in dimyristoylphosphatidylcholine vesicles using parinaric acid. , 1979, Biochemistry.

[33]  H. Mantsch,et al.  Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment. , 1993, Biochemistry.

[34]  A. Dunker,et al.  Proposed molten globule intermediates in fd phage penetration and assembly , 1991, FEBS letters.

[35]  W. Wickner.  Asymmetric orientation of a phage coat protein in cytoplasmic membrane of Escherichia coli. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[36]  G. Fasman,et al.  Computed circular dichroism spectra for the evaluation of protein conformation. , 1969, Biochemistry.

[37]  D. Marvin,et al.  Structure and assembly of filamentous bacterial viruses , 1975, Nature.

[38]  B. Sykes,et al.  Assignment of amide 1H and 15N NMR resonances in detergent-solubilized M13 coat protein: a model for the coat protein dimer. , 1992, Biochemistry.

[39]  A. Watts,et al.  Spin-label electron spin resonance study of bacteriophage M13 coat protein incorporation into mixed lipid bilayers. , 1987, Biochemistry.

[40]  R. Mitchell,et al.  Determination of protein secondary structure using factor analysis of infrared spectra. , 1990, Biochemistry.

[41]  H. D. de Jongh,et al.  ESR of spin-labeled bacteriophage M13 coat protein in mixed phospholipid bilayers. , 1990, Biochimica et biophysica acta.

[42]  I. Johnson,et al.  Environmental modulation of M13 coat protein tryptophan fluorescence dynamics. , 1989, Biochemistry.

[43]  G. Thomas,et al.  Structure similarity, difference and variability in the filamentous viruses fd, If1, IKe, Pf1 and Xf. Investigation by laser Raman spectroscopy. , 1983, Journal of molecular biology.

[44]  N. Kallenbach,et al.  The helix–coil transition in heterogeneous peptides with specific side‐chain interactions: Theory and comparison with CD spectral data , 1991, Biopolymers.

[45]  S. Opella,et al.  Protein backbone dynamics by solid-state and solution 15N NMR spectroscopy , 1987 .

[46]  A. Dunker,et al.  A NEW METHOD FOR DETERMINING PROTEIN SECONDARY STRUCTURE BY LASER RAMAN SPECTROSCOPY APPLIED TO fd PHAGE. , 1980, Biophysical journal.

[47]  W. Wickner.  Asymmetric orientation of phage M13 coat protein in Escherichia coli cytoplasmic membranes and in synthetic lipid vesicles. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[48]  A. Dunker,et al.  Circular dichroism studies of fd coat protein in membrane vesicles. , 1977, The Journal of biological chemistry.

[49]  D. Banner,et al.  Structure of the protein and DNA in fd filamentous bacterial virus , 1981, Nature.

[50]  A. Dunker,et al.  Raman spectroscopy and deuterium exchange of the filamentous phage fd. , 1984, Biochimica et biophysica acta.

[51]  S. Opella,et al.  Secondary structure of filamentous bacteriophage coat protein is preserved in lipid environments. , 1988, Journal of molecular biology.

[52]  B. Wallace,et al.  Differential absorption flattening optical effects are significant in the circular dichroism spectra of large membrane fragments. , 1987, Biochemistry.

[53]  P. Haris,et al.  A Fourier transform infrared investigation of the structural differences between ribonuclease A and ribonuclease S. , 1986, Biochimica et biophysica acta.

[54]  J. W. Parce,et al.  Lateral diffusion of M-13 coat protein in mixtures of phosphatidylcholine and cholesterol. , 1980, Biochemistry.

[55]  A. Visser,et al.  Time-resolved tryptophan fluorescence anisotropy investigation of bacteriophage M13 coat protein in micelles and mixed bilayers. , 1987, Biochemistry.

[56]  B. Sykes,et al.  Detergent-solubilized M13 coat protein exists as an asymmetric dimer. Observation of individual monomers by 15N, 13C and 1H nuclear magnetic resonance spectroscopy. , 1990, Journal of Molecular Biology.

[57]  P. Lu,et al.  Nitrogen-15 NMR spectroscopy of proteins in solution , 1990 .

[58]  B. Wallace,et al.  Circular dichroism analyses of membrane proteins: an examination of differential light scattering and absorption flattening effects in large membrane vesicles and membrane sheets. , 1984, Analytical biochemistry.