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.
Faculty of Dentistry, Kyushu University 61, Fukuoka, Japan.
H
H
Nagadome
K
K
Kawano
Y
Y
Terada
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