Adsorption of human lysozyme onto hydroxyapatite

FEBS Letters

Masaki, Kazuo¹  Kawano, Keiichi¹  Kamakura, Akito¹  Koganesawa, Nozomi¹  Terada, Yoshihiro²  Aizawa, Tomoyasu¹  Nitta, Katsutoshi¹  Matsuura, Atsushi¹  Nagadome, Hatsumi² 

[1] Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060, Japan;Faculty of Dentistry, Kyushu University, Fukuoka 812, Japan

关键词: Human lysozyme;    Hydroxyapatite;    High performance liquid chromatography;    Adsorption;   

DOI  :  10.1016/S0014-5793(97)01621-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To elucidate hydroxyapatite-protein interaction, mutant human lysozymes in which the surface charge was modified by site-directed mutagenesis were used. Five mutant human lysozymes (K1A, K13A, K33A, R10A, R14A) were expressed in yeast. The chromatographic behavior of these lysozymes was studied with a HPLC hydroxyapatite column. Elution molarities of K1A and R14A mutants were greatly lowered. While Lys-13 and Arg-10 are located around Lys-1 and Arg-14, K13A and R10A mutants bound onto hydroxyapatite stronger than K1A and R14A mutants. In combination with an X-ray crystal structure of human lysozyme, it is concluded that the adsorbing site of human lysozyme is at the back of the active site and that Arg-14, Lys-1, Arg-10 and Lys-13 play important roles in binding.

【 授权许可】

Unknown