First Crystal Structure of a Medicinally Relevant Gold Protein Complex: Unexpected Binding of
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Journal Article↗
Biomedical subjects
J Zou
Publications and source records attributed to J Zou.
Mitochondrial aconitase gene expression is regulated by testosterone and prolactin in prostate epithelial cells .
BACKGROUND: m-aconitase catalyzes the first step leading to the oxidation of citrate via the Krebs cycle. It is a constituitive enzyme in virtually all mammalian cells, found in excess, and is considered to be a regulatory or regulated enzyme. In contrast to these general relationships, prostate secretory epithelial cells possess a uniquely limiting mitochondrial (m-) aconitase which minimizes the oxidation of citrate. This permits the unique prostate function of accumulating and secreting extraordinarily high levels of citrate. Previous animal studies demonstrated that testosterone and prolactin regulate the level of m-aconitase specifically in citrate-producing prostate cells. The present studies were conducted to determine if testosterone and prolactin regulated the expression of the m-aconitase gene in prostate cells, and to determine the effect of the hormones on human prostate cells. METHODS: The studies were conducted with freshly prepared rat ventral, rat lateral, and pig prostate epithelial cells, and with the human malignant cell lines LNCaP and PC-3. The effects of 1 nM testosterone and 3 nM prolactin on the level of m-aconitase mRNA and on the transcription rate of m-aconitase were determined. RESULTS: The studies revealed that both prolactin and testosterone increase the levels of m-aconitase mRNA and the transcription rates of m-aconitase in rat ventral prostate cells, pig prostate cells, and human malignant prostate cells (LNCaP and PC-3). In contrast, both hormones decreased the level of m-aconitase mRNA and repressed m-aconitase gene transcription in rat lateral prostate cells. The hormonal regulation of m-aconitase corresponded with the levels of m-aconitase enzyme, m-aconitase activity, and citrate oxidation. CONCLUSIONS: In addition to the constitutive expression of m-aconitase, the m-aconitase gene is testosterone- and prolactin-regulated in specifically targeted prostate cells. The hormonal regulation of m-aconitase gene expression and biosynthesis of m-aconitase provide a regulatory mechanism for the oxidation of citrate, and consequently, the level of net citrate production by prostate. The hormonally increased expression and biosynthesis of m-aconitase in human malignant cells might be involved in the increased citrate oxidation associated with the development of true malignant cells in prostate cancer.
Aconitate Hydratase↗
Structure of Aspergillus niger epoxide hydrolase at 1.8 A resolution: implications for the structure and function of the mammalian microsomal class of epoxide hydrolases.
BACKGROUND: Epoxide hydrolases have important roles in the defense of cells against potentially harmful epoxides. Conversion of epoxides into less toxic and more easily excreted diols is a universally successful strategy. A number of microorganisms employ the same chemistry to process epoxides for use as carbon sources. RESULTS: The X-ray structure of the epoxide hydrolase from Aspergillus niger was determined at 3.5 A resolution using the multiwavelength anomalous dispersion (MAD) method, and then refined at 1.8 A resolution. There is a dimer consisting of two 44 kDa subunits in the asymmetric unit. Each subunit consists of an alpha/beta hydrolase fold, and a primarily helical lid over the active site. The dimer interface includes lid-lid interactions as well as contributions from an N-terminal meander. The active site contains a classical catalytic triad, and two tyrosines and a glutamic acid residue that are likely to assist in catalysis. CONCLUSIONS: The Aspergillus enzyme provides the first structure of an epoxide hydrolase with strong relationships to the most important enzyme of human epoxide metabolism, the microsomal epoxide hydrolase. Differences in active-site residues, especially in components that assist in epoxide ring opening and hydrolysis of the enzyme-substrate intermediate, might explain why the fungal enzyme attains the greater speeds necessary for an effective metabolic enzyme. The N-terminal domain that is characteristic of microsomal epoxide hydrolases corresponds to a meander that is critical for dimer formation in the Aspergillus enzyme.
Animals↗
Statistical theory of combinatorial libraries of folding proteins: energetic discrimination of a target structure.
A self-consistent theory is presented that can be used to estimate the number and composition of sequences satisfying a predetermined set of constraints. The theory is formulated so as to examine the features of sequences having a particular value of Delta=E(f)- (u), where E(f) is the energy of sequences when in a target structure and (u) is an average energy of non-target structures. The theory yields the probabilities w(i)(alpha) that each position i in the sequence is occupied by a particular monomer type alpha. The theory is applied to a simple lattice model of proteins. Excellent agreement is observed between the theory and the results of exact enumerations. The theory provides a quantitative framework for the design and interpretation of combinatorial experiments involving proteins, where a library of amino acid sequences is searched for sequences that fold to a desired structure.
Amino Acid Sequence↗
Transgenic mice overexpressing protein kinase C epsilon in their epidermis exhibit reduced papilloma burden but enhanced carcinoma formation after tumor promotion.
To determine the role that protein kinase C epsilon (PKCepsilon) may play in skin growth, differentiation, and tumor promotion, transgenic mice were generated that overexpressed an epitope-tagged protein kinase C epsilon (T7-PKCepsilon) in their epidermis using the human keratin 14 promoter. Three independent mouse lines that overexpressed the T7-PKCepsilon in their epidermis were produced. The three independent lines 206, 224, and 215 exhibited a 3-, 6-, and 18-fold elevation, respectively, in the level of PKCepsilon immunoreactive protein. Line 215 exhibited a 19-fold greater phosphatidylserine and 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated kinase activity than line 224. Line 206 exhibited a low basal T7-PKCepsilon activity, which failed to be stimulated by phosphatidylserine and TPA. All of the line 215 transgenic mice (F0 to the F2 generation) displayed phenotypic changes in the skin. The phenotypic changes progressed gradually, starting around 4-5 months of age, with mild dryness of the tail accompanied by hair loss and inflammation at the base of the tail. Hyperproliferation and ulceration of the affected regions were observed around 7-8 months of age. The hyperproliferative epidermis from the affected regions exhibited an expansion of the suprabasal epidermal cells. Inflammation and/or ulceration were also observed in the dorsal skin, the ears, and around the eyes. The line 215 mice, which expressed the highest level of PKCepsilon, were evaluated for sensitivity to mouse skin tumor promotion by TPA. Tumors were elicited by the initiation (7,12-dimethylbenz[a]anthracene, 100 nmol)-promotion (TPA, 5 nmol/twice weekly) protocol. The papilloma burden was reduced by 95-96% for male and female T7-PKCepsilon mice compared to wild-type controls. However, carcinomas developed rapidly in the T7-PKCepsilon mice treated with 7, 12-dimethylbenz[a]anthracene and TPA. These carcinomas appeared to form independently of prior papilloma development. These results demonstrate that PKCepsilon is an important regulator of skin tumor development.
9,10-Dimethyl-1,2-benzanthracene↗
Cloning, sequencing, and analysis of expression of a second IL-1beta gene in rainbow trout (Oncorhynchus mykiss).
The full-length sequence of a second IL-1beta gene (IL-1beta2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1beta gene, IL-1beta2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 3, which is 334 bp smaller in IL-1beta2. The transcript encoded by the IL-1beta2 gene contains a 5' untranslated region (UTR) of 121 bp, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation site 11 bp upstream of a 17-bp poly(A) tail. The predicted 254 amino acid sequence of the second IL-1beta gene has 82% similarity to the first gene, 45% similarity to carp IL-1beta, and 40% similarity to human IL-1beta. Comparison of the two trout genes reveals that the IL-1beta2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As with other nonmammalian IL-1beta genes, no interleukin-converting enzyme (ICE) cut site has been found but the alignment of the amino acid sequence with other species shows a possible cut site between Arg89 and Ala90 that would give arise to a 165-amino acid mature peptide. Expression studies performed by RT-PCR using primers specific for the IL-1beta2 transcript revealed a clear dose-dependent induction of this gene in cultured trout leukocytes by stimulation with lipopolysaccharide.
Amino Acid Sequence↗
Molecular cloning of the gene for interleukin-1beta from Xenopus laevis and analysis of expression in vivo and in vitro.
The Xenopus cDNA for interleukin-1beta (IL-1B) was cloned and sequenced. The gene contains 1,462 nucleotides that translate in a single reading frame to give a predicted 283-amino acid IL-1beta molecule. The translated molecule contains a single potential glycosylation site, a readily identifiable IL-1 family signature, and has highest homology to chicken IL-1beta by phylogenetic tree analysis and sequence similarity. It lacks a signal peptide in common with other known IL-1B genes, and lacks a clear ICE (caspase) cut site in common with other nonmammalian IL-1B genes sequenced to date. RT-PCR was used to study sites of IL-1B transcript expression, 24 h following injection of lipopolysaccharide (LPS). Expression was detected in the brain, liver, kidney, and spleen, with expression weakest in the brain and strongest in the spleen. No transcript expression was detectable following injection of saline. Northern blot analysis was used to quantify the induction of IL-1B expression in splenocytes following in vivo or in vitro stimulation with LPS. The results are discussed in relation to the potential role of IL-1beta in amphibian immune responses.
Amino Acid Sequence↗
Complexation of peptide with Cu2+ responsible to inducing and enhancing the formation of alpha-helix conformation.
Role of some metal ions on the conformations of peptides was examined by using a series of short alanine-based peptides with single Trp-His (W-H) interaction in different environments. Circular dichroism (CD), Trp (W) fluorescence emission, and Fourier transform infrared (FTIR) spectroscopy revealed that there is a conformational role of Cu2+ in inducing and enhancing the formation of alpha-helix conformation. The complexation of the peptide with Cu2+ is responsible to the conformational effect because the chelation is able to stabilize peptide with an alpha-helix conformation. The possible factors affecting the role of Cu2+ are discussed in the paper. The results in this paper are useful to understand the important structural role of Cu2+ in protein folding and the possible mechanism in some neurodegenerative diseases such as Alzheimer's disease.
Amino Acid Sequence↗
The pyruvate dehydrogenase E1 alpha gene is testosterone and prolactin regulated in prostate epithelial cells.
The prostate gland of humans and other animals has the unique function of accumulating and secreting extraordinarily high levels of citrate. The prostate secretory epithelial cells synthesize citrate which, due to a limiting mitochondrial (m-) aconitase, accumulates rather than being oxidized. Thus citrate is essentially an end product of metabolism in prostate. For continued net citrate production, a continual source of oxaloacetate (OAA) and acetyl CoA is required. Glucose via pyruvate oxidation provides the source of Acetyl CoA. In prostate cells, citrate production is regulated by testosterone and/or by prolactin. Both hormones selectively regulate the level and activity of pyruvate dehydrogenase E1 alpha (E1a) in animal prostate cells; thereby regulating the availability of acetyl CoA for citrate synthesis. Studies were conducted to determine if testosterone and prolactin might regulate the expression of the E1a gene in prostate epithelial cells. Prolactin treatment of rat ventral and lateral prostate cells and human PC3 cells increased the levels of E1a mRNA and the rates of transcription of the E1a gene. Testosterone also increased the mRNA level and transcription of E1a in rat ventral prostate cells, and in PC3 cells transfected with androgen receptor. However, testosterone treatment resulted in a repression of E1a gene expression in lateral prostate cells. Evidence is presented which supports the view that prolactin regulation of E1a is mediated via PKC. The rapidity of the effects of both hormones is representative of an immediate-early gene response. To our knowledge this represents the first report in any mammalian cells that, in addition to its constitutive expression in all mammalian cells, the E1a gene is a hormonally-regulated gene in specifically targeted prostate epithelial cells.
Animals↗
The plastidic phosphoglucomutase from Arabidopsis. A reversible enzyme reaction with an important role in metabolic control.
An Arabidopsis cDNA (AtPGMp) encoding the plastidic phosphoglucomutase (PGM) predicted a 623-amino acid protein with an N-terminal sequence typical of a plastid signal peptide. Expression of a recombinant protein in Escherichia coli confirmed its enzyme activity. The recombinant enzyme had an apparent K(m) value of 98.5 microM and a V(max) of 4.48 micromol min(-1) (mg protein)(-1). The Calvin cycle intermediates fructose-1,6-bisphosphate and ribulose-1, 5-bisphosphate exerted an inhibitory effect on PGM activity, supporting its proposed involvement in controlling photosynthetic carbon flow. A point mutation was identified in the AtPGMp gene of the Arabidopsis pgm-1 mutant. The mutation in the mutant transcript generated a stop codon at about one third of the wild-type open reading frame, and thus rendered the polypeptide nonfunctional. Storage lipid analysis of the pgm-1 mutant seeds showed a 40% reduction in oil content compared with that of wild type. Our results indicate that plastidic PGM is an important factor affecting carbon flux in triacylglycerol accumulation in oilseed plants, most likely through its essential role in starch synthesis.
Amino Acid Sequence↗
Cancer mortality in the high background radiation areas of Yangjiang, China during the period between 1979 and 1995.
The objective of the present study was to estimate cancer risk associated with the low-level radiation exposure of an average annual effective dose of 6.4 mSv (including internal exposure) in the high background-radiation areas (HBRA) in Yangjiang, China. The mortality survey consisted of two steps, i.e., the follow-up of cohort members and the ascertainment of causes of death. The cohort members in HBRA were divided into three dose-groups on the basis of environmental dose-rates per year. The mortality experiences of those three dose groups were compared with those in the residents of control areas by means of relative risk (RR). During the period 1987-1995, we observed 926,226 person-years by following up 106,517 subjects in the cohort study, and accumulated 5,161 deaths, among which 557 were from cancers. We did not observe an increase in cancer mortality in HBRA (RR = 0.96, 96% CI, 0.80 to 1.15). The combined data for the period 1979-95 included 125,079 subjects and accumulated 1,698,316 person-years, observed 10,415 total deaths and 1,003 cancer deaths. The relative risk of all cancers for whole HBRA as compared with the control area was estimated to be 0.99 (95% CI, 0.87 to 1.14). The relative risks of cancers of the stomach, colon, liver, lung, bone, female breast and thyroid within whole HBRA were less than one, while the risks for leukemia, cancers of the nasopharynx, esophagus, rectum, pancreas, skin, cervix uteri, brain and central nervous system, and malignant lymphoma were larger than one. None of them were significantly different from RR = 1. Neither homogeneity tests nor trend tests revealed any statistically significant relationship between cancer risk and radiation dose. We did not find any increased cancer risk associated with the high levels of natural radiation in HBRA. On the contrary, the mortality of all cancers in HBRA was generally lower than that in the control area, but not statistically significant.
Adolescent↗
Excess relative risk of solid cancer mortality after prolonged exposure to naturally occurring high background radiation in Yangjiang, China.
A study was made on cancer mortality in the high-background radiation areas of Yangjiang, China. Based on hamlet-specific environmental doses and sex- and age-specific occupancy factors, cumulative doses were calculated for each subject. In this article, we describe how the indirect estimation was made on individual dose and the methodology used to estimate radiation risk. Then, assuming a linear dose response relationship and using cancer mortality data for the period 1979-1995, we estimate the excess relative risk per Sievert for solid cancer to be -0.11 (95% CI, -0.67, 0.69). Also, we estimate the excess relative risks of four leading cancers in the study areas, i.e., cancers of the liver, nasopharynx, lung and stomach. In addition, we evaluate the effects of possible bias on our risk estimation.
Adolescent↗
A case-control study of nasopharyngeal carcinoma in the high background radiation areas of Yangjiang, China.
The main purposes of this study were to identify the major determinants of nasopharyngeal carcinoma (NPC) in the high-background radiation areas (HBRA) in Yangjiang, China and to evaluate their potential confounding effects on the NPC risk associated with exposure to high background radiation. A matched case-control study was conducted using those who died of NPC during the period 1987-1995. Two controls were randomly selected for each case from those who died from causes other than malignancies and external causes. Cases and their controls were matched with respect to sex and the years of birth and death (+/- 5 years). Study subjects' next-of-kin were interviewed using a standardized questionnaire to collect information on socioeconomic status, dietary habits, tobacco smoking and alcohol consumption, disease history, pesticide use, medical X-ray exposure, the family history of NPC and so on. We succeeded in interviewing 97 cases and 192 controls. Univariate conditional logistic regression analysis showed that NPC risk was associated with the consumption of salted fish, homemade pickles, and fermented soy beans, education levels, the history of chronic rhinitis, and the family history of NPC. Multivariate conditional logistic regression analysis revealed that education levels (Odds ratio (OR) for middle school or higher levels vs. no school education = 3.8, 95% CI = 1.2 to 11.8), salted fish intake (OR = 3.2, 95% CI = 1.7 to 6.1), the history of chronic rhinitis (OR = 3.6, 95% CI = 1.3 to 10.1), and the family history of NPC (OR = 14.2, 95% CI = 2.7 to 73.4) were independent risk factors of NPC. Tobacco smoking (OR = 1.2, 95% CI = 0.7 to 2.1), and alcohol consumption (OR = 0.9, 95% CI = 0.5 to 1.9) were not significantly related to NPC risk. The ORs of NPC risk comparing HBRA and a nearby control area before and after adjustment for the major risk determinants identified in the present study were 0.86 (95% CI = 0.50 to 1.50) and 0.87 (95% CI = 0.45 to 1.67), respectively. Salted fish intake was a strong risk factor of NPC. Education, the history of chronic rhinitis and the family history of NPC were also related to NPC risk. The exposure to high background radiation in HBRA of Yangjiang was not related to NPC risk with or without the adjustment for those major risk determinants of NPC.
Adult↗
Zinc induces mitochondria apoptogenesis in prostate cells.
BACKGROUND AND PURPOSE: Prostate secretory epithelial cells have the unique function and capability of accumulating extremely high intracellular levels of zinc. One of the effects of this accumulation is inhibition of cell growth due, in part, to an increase in apoptosis. The present studies were conducted to determine if this zinc-induced apoptosis involves stimulation of mitochondrial apoptogenesis. MATERIALS AND METHODS: The PC-3 a human malignant prostate cell line, which is zinc accumulating, was exposed to medium supplemented with physiologic levels of zinc. RESULTS: By 24 h, zinc treatment resulted in the translocation of cytochrome c from the mitochondria to the cytosol, the activation of caspase-9 and caspase-3, and eventually, the cleavage of nuclear poly(ADP)-ribose polymerase (PARP). Under similar conditions, exposure of freshly prepared rat ventral prostate cells (which are also zinc accumulating) resulted in increased apoptosis following translocation of cyochrome c and activation of caspases-9 and 3. The human prostate PZ-HPV-7 cells, which do not accumulate zinc, did not exhibit any apoptotic effect from zinc treatment. CONCLUSION: The accumulation of high intracellular levels of zinc by prostate cells induces mitochondrial apoptogenesis. This represents a newly identified physiological effect of zinc in the regulation of prostate cell growth.
Animals↗
[A randomized controlled study of ligustrazine in combination with propranolol for prevention of recurrent esophageal varices bleeding].
OBJECTIVE: To assess the efficacy of Ligustrazine in combination with propranolol in the prevention of recurrent esophageal varices bleeding following liver cirrhosis, and its act mechanism. METHODS: A prospective controlled study was conducted on 74 patients, in whom 38 belonged to treatment group, and 36 to control group. By detecting the portal system hemodynamics in patients with portal hypertension and esophageal varices using color Doppler ultrasound technique, the therapeutic efficacy and safety were investigated. Meanwhile the blood pressure, heart rate, hepatic and renal function were dynamically observed. RESULTS: After four weeks administration of the drugs, the flow of portal vein and splenic vein, the diameter of portal vein and splenic vein in the treatment group were significantly decreased with the values of 1152.36A387.46 ml/min, 529.35A326.31 ml/min, 1.36A0.28 cm, and 0.94A0.19 cm, respectively. No adverse effect was observed on the system circulation and the liver function. In the follow-up period of two years, the rebleeding rate and mortality rate in the placebo group were higher than that in the treatment group, but patients with liver cirrhosis Grade C in the two groups were not significant different. CONCLUSION: Low dose ligustrazine plus propranolol is a safe and effective therapy in preventing recurrent esophageal varices bleeding, and worth further trial.
Adult↗
Utility of the Arabidopsis FAE1 and yeast SLC1-1 genes for improvements in erucic acid and oil content in rapeseed.
High-erucic acid (HEA) Brassica napus cultivars are regaining interest in industrial contexts. Erucic acid and its derivatives are important renewable raw materials utilized in the manufacture of plastic films, in the synthesis of Nylon 13,13, and in the lubricant and emollient industries. Theoretically, the highest level of erucic acid that can be achieved by means of classical breeding is 66 mol%; however, using new approaches on the basis of genetic engineering, it might be possible to develop a B. napus cultivar containing levels of erucic acid significantly above 66 mol% (>80 mol%). In an attempt to increase the amounts of very-long-chain fatty acids (VLCFAs), and erucic acid in particular, in Canadian HEA B. napus cultivars, we have focused on two targets using a transgenic approach. We examined both the role/function of the Arabidopsis thaliana FAE1 (fatty acid elongase) gene by expressing it under the control of the seed-specific napin promoter in B. napus germplasm with analysis of the changes in VLCFA content in the seed oil of transgenic lines, and the performance of the yeast SLC1-1 (sphingolipid compensation mutant) in B. napus cv. Hero transgenic progeny in the field. Here, we report analyses of the contents of 22:1, total VLCFAand oil in the seed oil, as well as seed yield of the field-grown FAE1 and SLC1-1 B. napus cv. Hero progeny.
Acetyltransferases↗
Effects of resveratrol on oxidative modification of human low density lipoprotein.
OBJECTIVE: To determine the antioxidative effects of resveratrol (RES), a polyphenolic compound in red wine, on the oxidation of human low density lipoprotein (LDL) using two different oxidation systems. METHODS: Oxidation of LDL was induced by adding either Cu2+ or an azo compound. The extent of LDL modification was assessed by measuring the formation of thiobarbituric acid reactive substances (TBARS), the relative electrophoretic mobilities (REM), and the amount of oxidized LDL degradation by macrophages. RESULTS: During Cu(2+)-induced oxidation, RES reduced TBARS formation in LDL by 70.5%, REM of LDL by 42.3% and the amount of macrophage degradation by 65.7%, respectively. The lag phase of LDL oxidation was also delayed by adding RES both in the copper ion and azo compound-induced oxidation systems. CONCLUSION: RES can protect LDL against both Cu(2+)-induced and azo compound-initiated oxidative modification in vitro, which might be due to its free radical scavenging capacity.
Animals↗
[Automatic modulation of refractoriness of His-Purkinje system during atrioventricular nodal reentrant tachycardia].
OBJECTIVE: To illustrate the automatic modulation of refractoriness of His-Purkinje system during atrioventricular nodal reentrant tachycardia (AVNRT) and to discuss the possible mechanisms. METHODS: Programmed electrical stimulations were performed in high right atrium (HRA) in 8 patients with AVNRT before ablation to induce tachycardia and electrocardiagraphic recordings were done synchronically when AVNRT appeared. RESULTS: All the patients had 2:1 atrioventricular (A-V) conduction when AVNRT began, 2 of whom were blocked below His bundle, 5 above His bundle and 1 unclear. After a duration of 14.03 +/- 10.03 s of 2:1 A-V conduction, 1:1 A-V conduction with bundle banch block appeared, 3 of which were right bundle branch block (RBBB), 3 left bundle branch block (LBBB), and 2 with both. Bundle branch block disappeared after a duration of 6.87 +/- 11.26 s. CONCLUSION: Effective refractory period (ERP) of His-Purkinje system at the beginning of AVNRT was modulated automatically within less than 30-60 s and thus facilitated nodal-ventricular conduction. The mechanism of this is electrical remodeling.
Adult↗