Adsorption of human lysozyme onto hydroxyapatite. Identification of its adsorbing site using site-directed mutagenesis.

Abstract:

: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.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Aizawa T,Koganesawa N,Kamakura A,Masaki K,Matsuura A,Nagadome H,Terada Y,Kawano K,Nitta K

doi

10.1016/s0014-5793(97)01621-9

subject

Has Abstract

pub_date

1998-01-30 00:00:00

pages

175-8

issue

2

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(97)01621-9

journal_volume

422

pub_type

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