Glu375Gln and Asp225Val mutants: about the nature of the covalent linkages between heme group and apo-Protein in bovine lactoperoxidase

In analogy with studies previously reported for myeloperoxidase (Kooter, I. M.; Moguilevsky, N.; Bollen, A.; Van der Veen, L. A.; Otto, C.; Dekker, H. L.; Wever, R. J. Biol. Chem. 1999, 274, 26794), we examined for bovine lactoperoxidase the effect of mutation of Asp225 and Glu375, the residues thou...

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Published in:Bioorganic & medicinal chemistry letters 2001-11, Vol.11 (21), p.2827-2831
Main Authors: Suriano, Gianpaolo, Watanabe, Shikiko, Ghibaudi, Elena Maria, Bollen, Alex, Pia Ferrari, Rosa, Moguilevsky, Nicole
Format: Article
Language:English
Subjects:
Amino Acid Substitution
Analytical, structural and metabolic biochemistry
Animals
Aspartic Acid - chemistry
Base Sequence
Biological and medical sciences
Blotting, Western
Cattle
CHO Cells
Cricetinae
DNA Primers
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Glutamic Acid - chemistry
Glycine - chemistry
Heme - chemistry
Lactoperoxidase - chemistry
Mutation
Oxidoreductases
Valine - chemistry
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Summary:In analogy with studies previously reported for myeloperoxidase (Kooter, I. M.; Moguilevsky, N.; Bollen, A.; Van der Veen, L. A.; Otto, C.; Dekker, H. L.; Wever, R. J. Biol. Chem. 1999, 274, 26794), we examined for bovine lactoperoxidase the effect of mutation of Asp225 and Glu375, the residues thought to be responsible for the covalent binding of the heme group to the apoprotein. Starting from the plasmid encoding rbLPO (Watanabe, S.; Varsalona, F.; Yoo, Y.; Guillaume, J. P.; Bollen, A.; Shimazaki, K.; Moguilevsky, N. FEBS Letters 1998, 441, 476), which was engineered to carry mutations in correspondence of those residues, the mutants Asp225Val and Glu375Gln were expressed in CHO cells and their products purified and characterized. Unequivocal evidence about the existence of ester linkages as well as their relative contribution to the specific spectroscopic and catalytic properties of bLPO is here discussed. Unequivocal evidence about the existence of ester linkages as well as their relative contribution to the specific spectroscopic and catalytic properties of lactoperoxidase is here discussed.
ISSN:0960-894X
1464-3405
DOI:10.1016/S0960-894X(01)00533-9