Solution State Structure of P1, the Mimetic Peptide Derived from IgM Antigen Apo B-100 by NMR
1
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
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. Ex-perimental results show that P1 exhibit partial of β-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 0.33Å. The solution state P1 has mixed structure of β-sheet (Glu[1] to Cys[12]) and random coil (Pro[13] to Lys[20]). These NMR results are well consistent with secondary structure from ex-perimental 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 monoclo-nal antibodies.
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. Ex-perimental results show that P1 exhibit partial of β-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 0.33Å. The solution state P1 has mixed structure of β-sheet (Glu[1] to Cys[12]) and random coil (Pro[13] to Lys[20]). These NMR results are well consistent with secondary structure from ex-perimental 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 monoclo-nal antibodies.
참고문헌 (20)
- I. D. Cruzado, , J. Lipid Res, 39, pp.205~ (1998)
- R. W. Mahley, , J. Lipid Res, 25, pp.1277~ (1984)
- R. A. Davis, , Biochem. Lipid (1991)
- B. H. Browman, "Hepatic Plasma Proteins Mechanisms of Function and Regulations" (1993), Academic Press, Inc.
- V. N. Schumaker, , Adv. Protein, 45, pp.205~ (1994)
- S. H. Chen, , Biol. Chem, 261, pp.12918~ (1986)
- T. J. Knott, , Nature, 323, pp.734~ (1986)
- M. L. Phillips, , J. Lipid Res, 30, pp.415~ (1989)
- A. D. Cardin, , Biochem. Biophys. Acta, 877, pp.366~ (1986)
- G. C. Chen, , J. Biol. Chem, 264, pp.14369~ (1989)
- R. A. Davis, , J. Biol. Chem, 265, pp.10005~ (1990)
- G. C. Chen, , J. Biol. Chem, 264, pp.14369~ (1989)
- C. Ettelaie, , Biochem. Biophy. Acta, 1345, pp.237~ (1997)
- C. Y. Yang, , Nature, 323, pp.738~ (1986)
- H. Jamila, , J. Lipid Res, 39, pp.1448~ (1998)
- S. W. Law, , Proc. Nati. Acad. Sci. U. S. A, 83, pp.8142~ (1988)
- U. Stelnbrecher, , J. Biol. Chem, 262, pp.3603~ (1987)
- Sungran Lee, "Solution State Structure of pB1, the Mimotopic Peptide of Apolipoprotein B-100, by NMR", Bulletin of the Korean Chemical Society, 25(12), pp.1845~1849 (2004), 대한화학회
- 김대성, "Conformational Studies of Macrocyclic Corrin-Ring of Coenzyme B12 by NMR Methods", Journal of the Korean Magnetic Resonance Society, 3(1), pp.44~51 (1999), 한국자기공명학회
- N. J. Greenfield, , Trac-Trends Anal. Chem, 18, pp.236~ (1999)