Identification of catalytically important amino acids in human ceruloplasmin by site‐directed mutagenesis

FEBS Letters

Stenberg, Leisa M.¹  Brown, Mark A.¹  Mauk, A.Grant¹ 

[1] Department of Biochemistry and Molecular Biology, and the UBC Centre for Blood Research, Faculty of Medicine, University of British Columbia, 2146 Health Sciences Mall, Vancouver, BC, Canada V6T 1Z3

关键词: Ceruloplasmin;    Copper;    Ferroxidase;    Iron;    Diamine oxidase;    Site-directed mutagenesis;    CP;    ceruloplasmin;    ELISA;    enzyme-linked immunosorbent assay;    hCP;    human ceruloplasmin;    rhCP;    recombinant human ceruloplasmin;    TBS;    Tris-buffered saline;   

DOI  :  10.1016/S0014-5793(02)02652-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The involvement of amino acid residues previously proposed on the basis of structural data to have roles in the ferroxidase and diamine oxidase activities of human ceruloplasmin was investigated. Variants of human ceruloplasmin, in which residues proposed to be involved in electron transfer and/or iron-binding had been altered by site-directed mutagenesis, were expressed in HEK293 cells. E633A and E597A/H602A variants exhibited reduction in both activities by 50–60% compared to recombinant wild-type ceruloplasmin. The variant E935A/H940A had reduced ferroxidase activity (50%) but unaltered diamine oxidase activity, whereas the variant E971A exhibited enhanced diamine oxidase activity. For the L329M variant, both activities were identical to those of wild-type ceruloplasmin.

【 授权许可】

Unknown