Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Solution State Structure of P1, the Mimetic Peptide Derived from IgM Antigen Apo B-100 by NMR

  • Kim, Gilhoon (Department of Applied Chemistry, Hanyang University) ;
  • Lee, Hyuk (Department of Applied Chemistry, Hanyang University) ;
  • Oh, Hyewon (Department of Applied Chemistry, Hanyang University) ;
  • Won, Hoshik (Department of Applied Chemistry, Hanyang University)
  • Received : 2016.08.02
  • Accepted : 2016.08.23
  • Published : 2016.09.20
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Abstract

Apolipoprotein B-100 (Apo-B100) is a major component of low density lipoprotein (LDL). Apo B-100 protein has 4,536 amino acid sequence and these amino acids are classified into peptide groups A to G with subsequent 20 amino acids (P1-P302). The peptide groups were act as immunoglobulin (Ig) antigens which oxidized via malondialdehyde (MDA). The mimetic peptide P1 (EEEMLENVSLVCPKDAT RFK) out of D-group peptides carrying the highest value of IgG antigens were selected for structural studies that may provide antigen specificity. Circular Dichroism (CD) spectra were measured for peptide secondary structure in the range of 190-250 nm. Experimental results show that P1 exhibit partial of ${\beta}-sheet$ and random coil structure. Homonuclear (COSY, TOCSY, NOESY) 2D-NMR experiments were carried out for NMR signal assignments and structure determination for P1. On the basis of these completely assigned NMR spectra and distance data, distance geometry (DG) and Molecular dynamics (MD) were carried out to determine the structures of P1. The proposed structure was selected by comparisons between experimental NOE spectra and back calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P1 obtained upon superposition of all atoms was in the range $0.33{\AA}$. The solution state P1 has mixed structure of ${\beta}-sheet$ (Glu[1] to Cys[12]) and random coil (Pro[13] to Lys[20]). These NMR results are well consistent with secondary structure from experimental results of circular dichroism. Structural studies based on NMR may contribute to the studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.

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References

  1. I. D. Cruzado, S. L. Cockrill, C. J. McNeal, and R. D. Macfarlane, J. Lipid Res. 39, 205 (1998)
  2. R. W. Mahley, T. L. Innerarity, S. C. Jr. Rall, and K. H. Wesgraber, J. Lipid Res. 25, 1277 (1984)
  3. R. A. Davis, Biochem. Lipid (1991)
  4. B. H. Browman, Hepatic Plasma Proteins Mechanisms of Function and Regulations, Academic Press, Inc., London (1993)
  5. V. N. Schumaker, M. L. Phillips, and J. E. Chatterton, Adv. Protein 45, 205 (1994) https://doi.org/10.1016/S0065-3233(08)60641-5
  6. S. H. Chen, C. Y. Yang, P. F. Chen, D. Setzer, M. Tamimura, W. H. Li, A. M. Gotto, and L. J. Chan, Biol. Chem. 261, 12918 (1986)
  7. T. J.Knott, R. J.Pease, L. M.Powell, S. C.Wallis, S. C.Rall Jr, T. L. Innerarity, B. Blackhart, W. H. Taylor, Y. Marcel, R. Milne, D. Johnson, M. Fuller, A. J. Lusis, B. J. McCarthy, R. W. Mahley, B. L. Wilson, J. Scott, Nature 323, 734 (1986) https://doi.org/10.1038/323734a0
  8. M. L. Phillips, and V. N. Schumaker, J. Lipid Res. 30, 415 (1989)
  9. A. D. Cardin, and R. L. Jackson, Biochem. Biophys. Acta. 877, 366 (1986) https://doi.org/10.1016/0005-2760(86)90201-8
  10. G. C. Chen, S. Zhu, D. A. Hardman, J. W. Schilling, K. Lau, and J. P. Kane, J. Biol. Chem. 264, 14369 (1989)
  11. R. A. Davis, R. N. Thrift, C. C. Wu, and K. E. Howell, J. Biol. Chem. 265, 10005 (1990)
  12. G. C. Chen, S. Zhu, D. A. Hardman, J. W. Schilling, K. Lau, J. P. Kane, J. Biol. Chem., 264, 14369 (1989)
  13. C. Ettelaie, P. I. Haris, N. J. James, B. Wilbourn, J. M. Adam, and K. R. Bruckdorfer, Biochem. Biophy. Acta. 1345, 237 (1997) https://doi.org/10.1016/S0005-2760(96)00185-3
  14. C. Y. Yang, S. H. Chen, S. H. Gianturco, W. A. Bradley, J. T. Sparrow, M. Tanimura, W. H. Li, D. A. Sparrow, S. Deloof, M. Rosseneu, F. S. Lee, Z. W. Gu, A. M. Gotto, L. Chan, Nature 323, 738 (1986) https://doi.org/10.1038/323738a0
  15. H. Jamila, C. H. Chua, J. K. Dickson, Y. Chenb, M. Yana, S. A. Billerb, R. C. Gregga, J. R. Wetteraua, and D. A. Gordana, J. Lipid Res. 39, 1448 (1998)
  16. S. W. Law, S. M. Grant, and K. Hlguchi et al, Proc. Nati. Acad. Sci. U. S. A. 83, 8142 (1988)
  17. U. Stelnbrecher, J. Biol. Chem. 262, 3603 (1987)
  18. S. Lee, D. Kim, H. Kim, Y. Lee, H. Won, Bull. Korean Chem. Soc. 25, 12 (2004)
  19. D. Kim, J. Rho, and H. Won, J. Kor. Magn. Reson. Soc. 3, 44 (1999)
  20. N. J. Greenfield, Trac-Trends Anal. Chem. 18, 236 (1999) https://doi.org/10.1016/S0165-9936(98)00112-5