Horseradish peroxidase-dependent oxidation of deuteroporphyrin IX into chlorins

被引:13
作者
Dayan, FE
Duke, SO
Faibis, V
Jacobs, JM
Jacobs, NJ
机构
[1] Univ Mississippi, Sch Pharm, Natl Ctr Dev Nat Prod, NPURU,USDA,ARS, University, MS 38677 USA
[2] USDA ARS, So Weed Sci Lab, Stoneville, MS 38776 USA
[3] Dartmouth Coll, Sch Med, Dept Microbiol, Hanover, NH 03755 USA
基金
美国国家科学基金会;
关键词
chlorins; porphyrins; degradation; glutathione; peroxidase;
D O I
10.1006/abbi.1997.0533
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Chlorins are cyclic tetrapyrrole derivatives of great interest for use in photodynamic therapy, We have found that horseradish peroxidase (EC 1.11.1.7) (HRP) can convert deuteroporphyrin IX (Deutero) into chlorins, Some characteristics of this enzymatic transformation were investigated, The formation of chlorins was determined spectrophotometrically by monitoring the change in absorbance in the Q-band region (638 nn), The reaction occurred without addition of H2O2 and had a pH optimum of 7.5. The presence of thiol containing reductants, with a great preference for reduced glutathione, was required and could not be substituted by adding H2O2. Ascorbic acid acted as a potent inhibitor of the reaction, while other organic acids (citric and benzoic) had little to no inhibitory effect, The requirement for O-2 was suggested by the inhibitory effect of sodium hydrosulfite and was confirmed by carrying the assay in nitrogen-saturated solutions, Though the reaction occurred without adding H2O2, low amounts of H2O2 (3-30 mu M) were stimulatory to the assay, However, concentrations of 300 mu M H2O2 Or higher were inhibitory, Similarly, light was not required, but was stimulatory at low levels and inhibitory at high levels, Catalase and deferoxamine were inhibitory, but superoxide dismutase and mannitol had no effects. Kinetic analysis and respiratory studies suggest that HRP may initially react with reduced glutathione in a reaction that does not consume much oxygen, The ensuing steps, probably involving an oxygen free radical and porphyrin radical intermediates, consume a large amount of O-2 to oxidize Deutero into chlorin. (C) 1998 Academic Press.
引用
收藏
页码:27 / 34
页数:8
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