Molecular modeling of mammalian cytochrome P450s

The cytochrome P450s are a superfamily of hemoprotein enzymes responsible for the metabolism of a wide variety of xenobiotic and endogenous compounds. The individual P450s exhibit unique substrate specificity and stereoselectivity profiles which reflect corresponding differences in primary sequence and tertiary structure. In the absence of an experimental structure, models for mammalian P450s have been generated by their homology with bacterial P450s of known structure. The rather low sequence identity between target and template proteins renders P450 modeling a challenging task. However, the substrate recognition properties of several P450s are consistent with recently developed working models. This review summarizes the major concepts and current approaches of molecular modeling of P450s. Received 28 September 1999; received after revision 25 November 1999; accepted 31 December 1999     

Induction of cytochrome P 450 by phenobarbital in cats]

Phenobarbital administration to the Cat an animal hypersensitive to the actions of many toxic substances or to physiological compounds such as hormones, does not produce cytochrome P 450 induction. Consequently, hepatic storage of retinol remains unchanged. The specific and original characteristics of the Cat in this field are underlined by these results.     

Impact of neuropathic pain on the gene expression and activity of cytochrome P450side-chain-cleavage in sensory neural networks

Development of efficient therapy against chronic and stubborn pains requires fundamental identification of adequate cellular and molecular targets. This study combined cellular, molecular and biochemical approaches to investigate the gene expression and enzymatic activity of cytochrome P450side-chain-cleavage (P450scc) in spinal neural networks under normal and neuropathic pain states. P450scc is the key onset enzyme for steroidogenesis in endocrine glands and for neurosteroid biosynthesis in nerve cells. The P450scc gene was over-expressed in spinal and supra-spinal networks during neuropathic pain provoked by sciatic nerve ligature. Plasticity was observed in P450scc cellular distribution in pain circuits and its activity also increased inducing in vivo, hyper-secretion of pregnenolone and allopregnanolone which strongly stimulates type A receptors for g-aminobutyric acid, a pivotal neurotransmitter involved in pain modulation. These results, by establishing a direct link between neuropathic pain and neuroactive steroid formation in the nervous system, open new perspectives for chronic-pain modulation by endogenous neurosteroids.Received 8 June 2004; received after revision 2 July 2004; accepted 13 July 2004     

Joint effects of dietary protein levels and vitamin A additions on the induction of cytochrome P450 by DDT in rats]

A 5 P. 100 level of protein from casein in a diet does not allow vitamin A to modify significantly induction of cytochrome P 450 on the Rat receiving or not receiving DDT. When the protein increases to a 15 p. 100 level, the induction is better providing vitamin A is to be given. If protein and vitamin A are necessary for cytochrom P 450 induction, an increase of protein level remains inefficient without vitamin A.     

Effect of alpha-tocopherol (vitamin E) on the utilization of vitamin A and induction of cytochrome P 450 in rats receiving DDT]

During detoxication processes of DDT, there is, in the Rat, interference between vitamins A and E (D.1.alpha-tocopherol acetate). Vitamin E spares retinal and has a slighter but significant influence on P 450 cytochrome induction. These results are discussed.     

P2X4 and P2X6 receptors associate with VE-cadherin in human endothelial cells

We investigated the expression of P2X₄ and P2X₆ receptors on human umbilical vein endothelial cells (HUVECs) and found that both P2X receptor subtypes on plasma membranes are largely restricted to areas of cell-cell contact. Co-labelling experiments at the confocal and electron microscopy levels revealed that P2X₄ and P2X₆ receptors are strongly co-localised with the cell adhesion molecule VE-cadherin. The P2X₄ and P2X₆ receptors on plasma membranes at cellular junctions are rapidly (within 5 min) internalised specifically after decreasing extracellular [Ca²⁺]. Disruption of microfilaments, microtubules and integrin-mediated adhesion or stimulation of P2 receptors with ATP did not alter P2X₄ and P2X₆ receptor expression on HUVEC plasma membranes. Membraneous P2X₄ and P2X₆ receptors resisted extraction with Triton-X 100, whereas cytoplasmic P2X receptors were Triton-X 100 soluble. P2X₄ receptors, but not P2X₆ receptors, could be co-immunoprecipitated with VE-cadherin and vice versa. We conclude that P2X₄ and P2X₆ receptors are associated with VE-cadherin at HUVEC adherens junctions. Received 15 March 2002; revised 15 March 2002; accepted 19 March 2002     

Effect of lanthanons on substrate-induced difference spectra in rat liver microsomes

Summary Intravenously administered light lanthanons change spectral interactions in rat liver not only by decreasing the concentration of cytochrome P-450, but they also cause a qualitative change in the cytochrome P-450 molecule or its microenvironment.P. Arvela is a fellow of the Alexander-von-Humboldt-Stiftung.     

Influence du cadmium sur le métabolisme du calcium. Action sur le cytochrome P450

Summary Oral administration of cadmium to female rats for 6 weeks does not reduce the microsomal cytochrome P₄₅₀ levels in the liver and kidneys, nor the cytochrome P₄₅₀ content in the renal mitochondria.     

Cytochromes P450 and metabolism of xenobiotics

Cytochromes P450 (henceforth P450s) are involved in a variety of metabolic and biosynthetic processes. The number of known P450 enzymes exceeds 1000, while the endogenous substrates of most of them remain unknown. All P450 enzymes exhibit similarity in their structure and general mechanism of action; however, there are significant differences in the detailed function of individual enzymes as well as in the structures and properties of their active sites. This review discusses the properties of the most important P450 enzymes taking part in drug metabolism in humans. P450 3A4 is of paramount importance, because it is the most abundant P450 in the human liver and is known to metabolize the majority of drugs whose biotransformation is known. Genetically dependent variabilities of individual P450 activities and levels are described, documenting the importance of pharmacogenetics aimed at explaining differences in the response of the organism to various drugs. Received 7 November 2000; received after revision 9 January 2001; accepted 10 January 2001     

Effect of platinum derivatives on the inducible and repressible liver microsomal enzyme systems in the rat: inhibition of zoxazolamine-hydroxylase and induction of dimethyl-nitrosamine demethylase isoenzymes by cis-dichlorodiamine platinum (cis-PtCl2(NH3)2) and ammonium hexachloroplatinum (PtC16(NH4)2)]

Two platinum derivatives, cis-PtCl2(NH3)2 and PtCl6(NH4)2 have been studied for their effects on the Rat on cytochrome P450 in hepatic parenchyma on zoxazolamine-hydroxylase, a typical inducible system and on the two isoenzymes of dimethyl-nitrosamine demethylase, typical repressible systems. The inhibitory effect of PtCl6(NH4)2 on zoxazolamine-hydroxylase activity, previously shown by the authors, has been confirmed. The cis-PtCl2(NH3)2 also significantly inhibits zoxazolamine-hydroxylase activity. On the other hand, both of the platinum derivatives decrease cytochrome P450 level and enhance the dimethyl-nitrosamine metabolism. These various effects and their relationship are discussed.     

Effect of topically applied phenobarbital on O-dealkylase activity in mouse skin

Summary Topical application of phenobarbital to mice produces an increase in cutaneous microsomal proteins and apparently also of cytochrome P450, but results in a kinetically noncompetitive inhibition of p-nitrophenethole O-dealkylase activity in preparations of 10,000×g supernatant from skin.     

Nicotinamide administration alters the activities of hepatic microsomal mixed function oxidases

Summary Systemic action of nicotinamide significantly alters the activities of hepatic drug metabolizing enzymes. Male rats injected with nicotinamide have reduced levels of cytochrome P450, demethylases and aniline hydroxylase. The changes appear to be sex-dependent since in the case of female rats activities of p-nitroanisole-o-demethylase and aniline hydroxylase are enhanced whereas cytochrome P450 content remains unaltered.Authors gratefully acknowledge the encouragement given by Professor A.S. Paintal during the course of this investigation. One of us (J.K.B.) is grateful to the Council of Scientific and Industrial Research, New Delhi, for the award of a research fellowship.     

Stimulation of pituitary LH and FSH secretion after immunoneutralization of endogenous substance P in the cycling rat]

In the cycling female Rat, administration of substance P antisera performed at noon on the proestrous day provokes a significant increase in LH and FSH serum level, right before or after the preovulatory surges, but had no effect on its occurrence or magnitude. These results suggest that substance P has a normal physiological inhibitory role in gonadotropin regulation.     

P2P系统中基于双信任信息的信誉度改进模型

针对P2P网络中诸如诋毁、合谋、欺骗等安全性问题,提出一种新的基于双信任信息的分布式信任模型。区别对待节点提供服务和发送评价的能力,对节点拥有的资源进行分类;通过迭代求解,为每个节点的每类资源分别计算全局服务信任值和回馈信任值;为了防止恶意节点反复实施恶意行为,引入了惩罚机制,激励节点诚信交易。仿真实验表明,该模型能够迅速降低恶意节点的全局信誉值和恶意交易概率,降低了对无过失节点的不公平性。     

Vitamin A consumption in pregnant and non pregnant rats, and in castrated rats receiving or not receiving estradiol or pregnenolone; relation to tocopherol and cytochrome P-450]

Consumption of vitamin A and cytochrome P 450 by pregnant Rat is more important than in non pregnant and ovariectomised Rat. Estradiol implant in ovariectomised female has some, but slighter, influence only on retinol. These physiological situations have no action on the hepatic levels of tocopherols (total and alpha). These results are discussed.     

Interaction and interrelation of P2X7 and P2X4 receptor complexes in mouse lung epithelial cells

P2X4 and P2X7 receptors are ATP-gated ion channels that are co-expressed in alveolar epithelial type I cells. Both receptors are localized to the plasma membrane and partly associated with lipid rafts. Here we report on our study in an alveolar epithelial cell line of the molecular organization of P2X7R and P2X4R receptors and the effect of their knockdown. Native gel electrophoresis reveals three P2X7R complexes of ~430, ~580 and ~760 kDa. The latter two correspond exactly in size to signals of Cav-1, the structural protein of caveolae. Interestingly knockdown of P2rx7 affects protein levels, the intracellular distribution and the supramolecular organization of Cav-1 as well as of P2X4R, which is mainly detected in a complex of ~430 kDa. Our data suggest upregulation of P2X4R as a compensatory mechanism of P2X7R depletion.     

The effect of hypoxia on hepatic cytochromes and heme turnover in rats in vivo

We evaluated the effect of hypoxia (7% v/v) on hepatic heme turnover in vivo and microsomal heme protein content in male Sprague-Dawley rats. Hepatic heme protein turnover, measured as 14CO-production during continuous infusion of 5-14C-aminolevulinic acid, a precursor of nonerythrogenic heme, was decreased 60% during hypoxia and returned to control levels promptly after reoxygenation. Hepatic cytochrome P-450 content was decreased in hypoxic and 24-h reoxygenated animals. We conclude that normobaric hypoxia decreases hepatic cytochrome P-450 which could contribute to decreased drug metabolism in hypoxia. This decrease is probably due to heme oxygenase-independent breakdown of hepatic heme.     

公钥体系的混合式P2P信任模型

设计公钥体系的混合式P2P网络信任模型。采用层次化管理体系对节点进行认证,公钥体系保障节点间通信的安全性。通过局部信任、推荐信任、全局信任综合考察节点的可信情况。给出了模型的数学实现方法。仿真结果表明,模型能够有效提高节点的交易成功率。     

Hydrogen peroxide and hydroxyl radical involvement in the activation of caspase-3 in chemically induced apoptosis of HL-60 cells

Apoptosis of HL-60 cells induced by actinomycin D, H7, or daunorubicin was shown to involve the activation of caspase-3-like protease, 2 h after the addition of these drugs, based on microassay of enzyme activity by high-performance liquid chromatography. Catalase and a spin trap, N-t-butyl--phenylnitrone, which effectively inhibited the apoptosis induced by these drugs, also inhibited the activation of caspase-3-like protease. These results suggest that hydrogen peroxide and the hydroxyl radical are common mediators of caspase-3 activation caused by these chemicals, with apparently different functional mechanisms. Based on mitochondrial activity determined by oxygen consumption, complexes I, II, and IV were inhibited by actinomycin D. H7 inhibited complexes I and IV, 1 and 1.5 h respectively, after the addition of the drug to HL-60 cells. Daunorubicin inhibited complex IV, 1.5 h after the addition of the drug to HL-60 cells. Inhibition of complex IV by actinomycin D, H7, and daunorubicin were almost fully restored by the addition of cytochrome c. The release to the cytosol of cytochrome c by these drugs was also demonstrated by Western blot analysis. Addition of catalase inhibited the depression of complex IV activity induced by actinomycin D and H7. These observations indicate a direct relationship between hydrogen peroxide and the release of cytochrome c during apoptosis caused by actinomycin D, H7, and daunorubicin. Received 24 November 2000; received after revision 2 January 2001; accepted 30 January 2001