Recombinant bovine lactoperoxidase as a tool to study the heme environment in mammalian peroxidases

FEBS Letters

Yoo, Yung-Choon³  Watanabe, Shikiko¹  Bollen, Alex²  Shimazaki, Keiichi¹  Guillaume, Jean-Paul²  Moguilevsky, Nicole²  Varsalona, Franca² 

[1] Dairy Science Laboratory, Faculty of Agriculture, Hokkaido University, Sapporo, Japan;Applied Genetics, Faculty of Sciences, Université Libre de Bruxelles, 24 rue de l'Industrie, B-1400 Nivelles, Belgium;Institute of Immunological Science, Hokkaido University, Sapporo, Japan

关键词: Recombinant lactoperoxidase;    Myeloperoxidase;    Heme;    Peroxidase activity;    Chlorination;    Site-directed mutagenesis;    r-;    recombinant;    b-;    bovine;    LPO;    lactoperoxidase;    h-;    human;    MPO;    myeloperoxidase;    EPO;    eosinophil peroxidase;    TPO;    thyroid peroxidase;    CHO;    Chinese hamster ovary;    OPD;    O-phenylenediamine;    ABTS;    2;    2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid);    ELISA;    enzyme linked immunosorbent assay;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/S0014-5793(98)01595-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNA encoding bovine lactoperoxidase (LPO) has been expressed in CHO cells. The recombinant LPO was secreted as an enzymatically active single chain molecule presenting two immunoreactive forms of 88 kDa and 82 kDa, differing by their glycosylation. rLPO exhibited the characteristic absorbance spectrum with a Soret peak at 413 nm. Engineering of rLPO into a myeloperoxidase (MPO)-like molecule was attempted by substituting Gln-376 by Met, a residue known to achieve covalent binding with the heme in MPO. However, the resulting bovine LPO mutant failed to acquire the peculiar absorbance spectrum and the chlorinating activity of MPO, underlining the complex nature of interactions in the heme vicinity.

【 授权许可】

Unknown