Identification of the adsorbing site of lysozyme onto the hydroxyapatite surface using hydrogen exchange and 1H NMR.
The lysozyme-hydroxyapatite interaction was studied by measuring individual hydrogen-deuterium (H-D) exchange rates of amide protons. The H-D exchange reaction was initiated by transferring the lysozyme adsorbed on hydroxyapatite powder from H2O into D2O. After various H-D exchange time periods (pH 7.0, 25 degrees C), the complex was dissociated and the remaining hydrogen label was determined by 2D NMR analysis. The H-D exchange rate of amide protons of residues 9, 11, 13, and 83 was slowed in the hydroxyapatite-lysozyme complex compared with free lysozyme. Residues 9, 11 and 13 positioned at the back of the active site would be the location of the binding site.
Nagadome
H
H
Faculty of Dentistry, Kyushu University 61, Fukuoka, Japan.
Kawano
K
K
Terada
Y
Y
D000327:Adsorption; D000818:Animals; D002645:Chickens; D017886:Durapatite; D004529:Egg White; D006859:Hydrogen / Q000737:chemistry*; D006882:Hydroxyapatites / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009113:Muramidase / Q000737:chemistry*; D011487:Protein Conformation
D006882:Hydroxyapatites; D006859:Hydrogen; D017886:Durapatite; D009113:Muramidase