Abstract：
:In order to develop in vitro methods as an alternative to P450 animal testing in the drug discovery process, two main requisites are necessary: 1) gathering of data on animal homologues of the human P450 enzymes, currently very limited, and 2) bypassing the requirement for both the P450 reductase and the expensive cofactor NADPH. In this work, P450 2C20 from Macaca fascicularis, homologue of the human P450 2C8 has been taken as a model system to develop such an alternative in vitro method by two different approaches. In the first approach called "molecular Lego", a soluble self-sufficient chimera was generated by fusing the P450 2C20 domain with the reductase domain of cytochrome P450 BM3 from Bacillus megaterium (P450 2C20/BMR). In the second approach, the need for the redox partner and also NADPH were both obviated by the direct immobilization of the P450 2C20 on glassy carbon and gold electrodes. Both systems were then compared to those obtained from the reconstituted P450 2C20 monooxygenase in presence of the human P450 reductase and NADPH using paclitaxel and amodiaquine, two typical drug substrates of the human P450 2C8. The K(M) values calculated for the 2C20 and 2C20/BMR in solution and for 2C20 immobilized on electrodes modified with gold nanoparticles were 1.9 ± 0.2, 5.9 ± 2.3, 3.0 ± 0.5 μM for paclitaxel and 1.2 ± 0.2, 1.6±0.2 and 1.4 ± 0.2 μM for amodiaquine, respectively. The data obtained not only show that the engineering of M. fascicularis did not affect its catalytic properties but also are consistent with K(M) values measured for the microsomal human P450 2C8 and therefore show the feasibility of developing alternative in vitro animal tests.
journal_name
J Inorg Biochemjournal_title
Journal of inorganic biochemistryauthors
Rua F,Sadeghi SJ,Castrignanò S,Di Nardo G,Gilardi Gdoi
10.1016/j.jinorgbio.2012.05.017subject
Has Abstractpub_date
2012-12-01 00:00:00pages
277-84eissn
0162-0134issn
1873-3344pii
S0162-0134(12)00185-7journal_volume
117pub_type
杂志文章abstract：:Citrate occurs at about 0.1 mM in blood plasma and is the most likely small molecule plasma binder of both Al3+ and Fe3+. This paper assesses published stability constants for citrate binding to each metal ion. From pH 2 to 5 Al3+ forms a neutral complex with citrate that may be absorbed into the body in the upper reg...
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journal_title：Journal of inorganic biochemistry
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journal_title：Journal of inorganic biochemistry
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2011.04.004
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2010.01.001
更新日期：2010-04-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2006.09.017
更新日期：2006-12-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2015.05.010
更新日期：2015-09-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/0162-0134(88)80013-8
更新日期：1988-01-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2012.04.002
更新日期：2012-08-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/0162-0134(88)85025-6
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
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pub_type： 杂志文章
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2006.11.009
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
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pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2003.09.011
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journal_title：Journal of inorganic biochemistry
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
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abstract：:A number of 22 lanthanide-containing coordination compounds of sulfonamides 2 and 3 were prepared and characterized by elemental analysis and spectroscopic data. The new compounds proved to be highly potent inhibitors of isozymes I and II of carbonic anhydrase. ...
journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/0162-0134(93)85019-5
更新日期：1993-02-01 00:00:00
abstract：:The synthesis, physico-chemical characterization and cytotoxicity against five human tumoral cell lines (THP-1, U937, Molt-4, Colo205 and H460) of three new cobalt(II) coordination compounds are reported (i.e. Co(HL1)Cl (1), Co(HL2)Cl (2) and [Co(HL3)Cl]0.0.5 (CH3)2CHOH (3)). H2L2 (2-{[[2-hydroxy-3-(1-naphthyloxy)prop...
journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2016.05.003
更新日期：2016-08-01 00:00:00
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journal_title：Journal of inorganic biochemistry
pub_type： 杂志文章
doi：10.1016/j.jinorgbio.2007.03.002
更新日期：2007-06-01 00:00:00
