Molecular design and modeling of protein-heparin interactions
The methods and approaches taken to investigate heparin-apoE peptide interactions have involved a series of steps, including (1) identification of the heparin-binding domains of apoE, (2) determination of the minimal amino acid sequence regions involved in heparin binding, heparin-induced conformati...
Saved in:
Published in | Methods in enzymology Vol. 203; pp. 556 - 583 |
---|---|
Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
1991
|
Subjects | |
Online Access | Get more information |
Cover
Loading…
Summary: | The methods and approaches taken to investigate heparin-apoE peptide interactions have involved a series of steps, including (1) identification of the heparin-binding domains of apoE, (2) determination of the minimal amino acid sequence regions involved in heparin binding, heparin-induced conformational changes, and stability of apoE peptide structures in solution, (3) modeling of these peptide and oligosaccharide structures, and (4) examination of their behavior during molecular dynamics calculations to determine if the modeled complexes simulate the results of the solution study. The heparin-binding regions of apoE were determined by fragmentation of the protein and identification of the heparin-binding fragments by ligand-blotting procedures using 125I-labeled heparin. Studies with synthetic peptide fragments of various lengths and dot-blot procedures with 125I-labeled heparin identified the minimal residues critical for heparin-binding and CD studies established the prominent secondary structures of these domains. These studies also showed that heparin binds to the apoE(211-243) and apoE(129-169) regions to induce and stabilize beta-strand and alpha-helical peptide conformations. Secondary structure algorithms were used to identify the specific residues with the highest probabilities of forming alpha-helix and beta-strand structures. Based on the predictive algorithms, the apoE(211-234) and apoE(129-159) structures were built using the Insight program and their molecular interactions with various heparin oligosaccharide models were investigated by molecular dynamics. In agreement with the solution studies in the presence of salt, the molecular dynamics studies showed that the oligosaccharides stabilized the beta-strand and alpha-helical peptide configurations against simulated thermal denaturations. Further modeling studies are in progress to examine the mechanism of the heparin-induced increase in ordered structure of these peptides. |
---|---|
ISSN: | 0076-6879 1557-7988 |
DOI: | 10.1016/0076-6879(91)03030-K |