scholarly journals Induction of human spermine oxidase SMO(PAOh1) is regulated at the levels of new mRNA synthesis, mRNA stabilization and newly synthesized protein

2005 ◽  
Vol 386 (3) ◽  
pp. 543-547 ◽  
Author(s):  
Yanlin WANG ◽  
Amy HACKER ◽  
Tracy MURRAY-STEWART ◽  
Jennifer G. FLEISCHER ◽  
Patrick M. WOSTER ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Half Life ◽  
Cellular Response ◽  
Actinomycin D ◽  
Tumour Response ◽  
Spermine Oxidase ◽  
Direct Role ◽  
Mrna Stabilization ◽  
Polyamine Analogues ◽  
And Control

The oxidation of polyamines induced by antitumour polyamine analogues has been associated with tumour response to specific agents. The human spermine oxidase, SMO(PAOh1), is one enzyme that may play a direct role in the cellular response to the antitumour polyamine analogues. In the present study, the induction of SMO(PAOh1) enzyme activity by CPENSpm [N1-ethyl-N11-(cyclopropyl)methyl-4,8,diazaundecane] is demonstrated to be a result of newly synthesized mRNA and protein. Inhibition of new RNA synthesis by actinomycin D inhibits both the appearance of SMO(PAOh1) mRNA and enzyme activity. Similarly, inhibition of newly synthesized protein with cycloheximide prevents analogue-induced enzyme activity. Half-life determinations indicate that stabilization of SMO(PAOh1) protein does not play a significant role in analogue-induced activity. However, half-life experiments using actinomycin D indicate that CPENSpm treatment not only increases mRNA expression, but also leads to a significant increase in mRNA half-life (17.1 and 8.8 h for CPENSpm-treated cells and control respectively). Using reporter constructs encompassing the SMO(PAOh1) promoter region, a 30–90% increase in transcription is observed after exposure to CPENSpm. The present results are consistent with the hypothesis that analogue-induced expression of SMO(PAOh1) is a result of increased transcription and stabilization of SMO(PAOh1) mRNA, leading to increased protein production and enzyme activity. These data indicate that the major level of control of SMO(PAOh1) expression in response to polyamine analogues exposure is at the level of mRNA.

scholarly journals Spermine synthesis in the uterus of the ovariectomized rat in response to oestradiol-17β

1972 ◽  
Vol 128 (5) ◽  
pp. 1109-1115 ◽  
Author(s):  
D H. Russell ◽  
J J. Potyraj
Keyword(s):  
Enzyme Activity ◽  
Half Life ◽  
Actinomycin D ◽  
Deae Cellulose ◽  
Ovariectomized Rat ◽  
Enzymic Synthesis ◽  
Hormone Administration

We reported that spermidine and spermine pools in the uterus both doubled within 24h after oestradiol administration to castrated rats (Russell & Taylor, 1971). Now we have studied the enzymic synthesis of spermine (by spermidine-dependent S-adenosyl-l-methionine decarboxylase) and find that the activity of the enzyme(s) involved is elevated soon after hormone administration. Enzyme activity is increased within 4h and is five times that of controls within 24h. Cycloheximide or actinomycin D administered at the time of oestradiol injection completely blocked the increase in enzyme activity. The enzyme involved in spermine synthesis, S-adenosyl-l-methionine decarboxylase, with S-adenosyl-l-methionine and spermidine as required substrates, was partially purified on Sephadex and DEAE-cellulose columns. The decarboxylation of S-adenosyl-l-methionine could not be separated from the transfer of a propylamine moiety from the decarboxylated S-adenosyl-l-methionine to spermidine to form spermine. We were unable also to separate this system from the enzyme that formed spermidine when S-adenosyl-l-methionine and putrescine are used as substrates. Spermidine-stimulated S-adenosyl-l-methionine decarboxylase has an apparent half-life of 60min, identical with the half-life reported for putrescine-stimulated S-adenosyl-l-methionine decarboxylase. These results strongly suggest that the same enzyme(s) operate in the synthesis of both spermidine and spermine.

The role of polyamine catabolism in anti-tumour drug response

2003 ◽  
Vol 31 (2) ◽  
pp. 361-365 ◽  
Author(s):  
R.A. Casero ◽  
Y. Wang ◽  
T.M. Stewart ◽  
W. Devereux ◽  
A. Hacker ◽  
...  
Keyword(s):  
Cellular Response ◽  
Drug Response ◽  
Molecular Mechanisms ◽  
Oxidase Inhibitor ◽  
Polyamine Oxidase ◽  
Direct Role ◽  
Polyamine Catabolism ◽  
Polyamine Analogues ◽  
Cytotoxic Response ◽  
High Induction

Interest in polyamine catabolism has increased since it has been directly associated with the cytotoxic response of multiple tumour types to exposure to specific anti-tumour polyamine analogues. Human polyamine catabolism was considered to be a two-step pathway regulated by the rate-limiting enzyme spermidine/spermine N1-acetyltransferase (SSAT) that provides substrate for an acetylpolyamine oxidase (APAO). Further, the super-induction of SSAT by several anti-tumour polyamine analogues has been implicated in the cytotoxic response of specific solid-tumour phenotypes to these agents. This high induction of SSAT has been correlated with cellular response to the anti-tumour polyamine analogues in several systems and considerable progress has been made in understanding the molecular mechanisms that regulate the analogue-induced expression of SSAT. A polyamine response element has been identified and the transacting transcription factors that bind and stimulate transcription of SSAT have been cloned and characterized. The link between SSAT activity and cellular toxicity is thought to be based on the production of H2O2 by the activity of the constitutive APAO that uses the SSAT-produced acetylated polyamines. The high induction of SSAT and the subsequent activity of APAO are linked to the cytotoxic response of some tumour cell types to specific polyamine analogues. However, we have recently cloned a variably spliced human polyamine oxidase (PAOh1) that is inducible by specific polyamine analogues, efficiently uses unacetylated spermine as a substrate, and also produces toxic H2O2 as a product. The results of studies with PAOh1 suggest that it is an additional enzyme in polyamine catabolism that has the potential to significantly contribute to polyamine homoeostasis and drug response. Most importantly, PAOh1 is induced by specific polyamine analogues in a tumour-phenotype-specific manner in cell lines representative of the major forms of solid tumours, including lung, breast, colon and prostate. The sensitivity to these anti-tumour polyamine analogues can be significantly reduced if the tumour cells are co-treated with 250 μM of the polyamine oxidase inhibitor N1, N4-bis(2,3-butadienyl)-1,4-butanediamine (MDL 72,527), suggesting that the H2O2 produced by PAOh1 does in fact play a direct role in the observed cytotoxicity. These results strongly implicate PAOh1 as a new target that, in combination with SSAT, may be exploited for therapeutic advantage. The current understanding of the role and regulation of these two important polyamine catabolic enzymes are discussed.

SIALIC ACID IN THE PRIMARY AND DIFFERENTIAL DIAGNOSIS OF LUNG CANCER

2017 ◽  
Vol 63 (6) ◽  
pp. 926-932
Author(s):  
Lyudmila Belskaya ◽  
Viktor Kosenok ◽  
Ж. Массард
Keyword(s):  
Lung Cancer ◽  
Enzyme Activity ◽  
Optimization Problems ◽  
Primary Lung Cancer ◽  
Nonparametric Test ◽  
Nonsmall Cell Lung Cancer ◽  
Group Differences ◽  
Diagnosis Of Lung Cancer ◽  
And Control ◽  
Control Study

So far optimization problems for diagnostics and prognostication aids remained relevant for lung cancer as a leader in the structure of cancers. Objective: a search for regularities of changes in the saliva enzyme activity in patients with nonsmall cell lung cancer. In the case-control study, 505 people took part, divided into 2 groups: primary (lung cancer, n=290) and control (conventionally healthy, n=215). All the participants went through a questionnaire survey, saliva biochemical counts, and a histological verification of their diagnosis. The enzyme activity was measured with spectrophotometry. Between-group differences were measured with the nonparametric test. It was shown that in terms of lung cancer, we observe metabolic changes, described with the decreased de Ritis coefficient (p

scholarly journals Biphasic effect of acute ethanol administration on rat liver tyrosine-2-oxoglutarate aminotransferase activity

1980 ◽  
Vol 186 (3) ◽  
pp. 755-761 ◽  
Author(s):  
A A B Badawy ◽  
B M Snape ◽  
M Evans
Keyword(s):  
Enzyme Activity ◽  
Rat Liver ◽  
Actinomycin D ◽  
Tyrosine Aminotransferase ◽  
Ethanol Metabolism ◽  
Ethanol Administration ◽  
Aminotransferase Activity ◽  
Biphasic Effect ◽  
Initial Decrease ◽  
Acute Ethanol

1. Acute ethanol administration causes a biphasic change in rat liver tyrosine aminotransferase activity. 2. The initial decrease is significant with a 200 mg/kg dose of ethanol, is prevented by adrenoceptor-blocking agnets and by reserpine, but not by inhibitors of ethanol metabolism, and exhibits many of the characteristics of the inhibition caused by noradrenaline. 3. The subsequent enhancement of the enzyme activity by ethanol is not associated with stabilization of the enzyme, but is sensitive to actinomycin D and cycloheximide. 4. It is suggested that the initial decrease in aminotransferase activity is caused by the release of catecholamines, whereas the subsequent enhancement may be related to the release of glucocorticoids.

scholarly journals Identification and characterization of a 44 kDa protein that binds specifically to the 3′-untranslated region of CYP2a5 mRNA: inducibility, subcellular distribution and possible role in mRNA stabilization

1996 ◽  
Vol 313 (3) ◽  
pp. 1029-1037 ◽  
Author(s):  
Olivier GENESTE ◽  
Françoise RAFFALLI ◽  
Matti A. LANG
Keyword(s):  
Half Life ◽  
Untranslated Region ◽  
Translational Regulation ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Blocking Agent ◽  
Subcellular Fractionation ◽  
Mrna Stabilization ◽  
Nuclear Extracts

Stabilization of mRNA is important in the regulation of CYP2a5 expression but the factors involved in the process are not known [Aida and Negishi (1991) Biochemistry 30, 8041–8045]. In this paper, we describe, for the first time, a protein that binds specifically to the 3′-untranslated region of CYP2a5 mRNA and which is inducible by pyrazole, a compound known to increase the half-life of CYP2a5 mRNA. We also demonstrate that pyrazole treatment causes an elongation of the CYP2a5 mRNA poly(A) tail, and that phenobarbital, which is transcriptional activator of the CYP2a5 gene that does not affect the mRNA half-life, neither induces the RNA-binding protein nor affects the poly(A) tail size. SDS/PAGE of the UV-cross-linked RNA–protein complex demonstrated that the RNA-binding protein has an apparent molecular mass of 44 kDa. The protein-binding site was localized to a 70-nucleotide region between bases 1585 and 1655. Treatment of cytoplasmic extracts with an SH-oxidizing agent, diamide, an SH-blocking agent, N-ethylmaleimide or potato acid phosphatase abolished complex-formation, suggesting that the CYP2a5 mRNA-binding protein is subject to post-translational regulation. Subcellular fractionation showed that the 44 kDa protein is present in polyribosomes and nuclei, and that its apparent induction is much stronger in polyribosomes than in nuclear extracts. We propose that this 44 kDA RNA-binding protein is involved in the stabilization of CYP2a5 mRNA by controlling the length of the poly(A) tail.

EFFECTS OF CRYSTAL SIZE AND ORIENTATION OF SUBSTRATES ON CELL ADHESION: IMPLICATION FOR MEDICAL IMPLANTS

2008 ◽  
Vol 22 (18n19) ◽  
pp. 3069-3081 ◽  
Author(s):  
SHAHAB FAGHIHI ◽  
HOJATOLLAH VALI ◽  
MARYAM TABRIZIAN
Keyword(s):  
Cellular Response ◽  
High Pressure Torsion ◽  
Control Cell ◽  
Cell Activity ◽  
Titanium Implants ◽  
Polycrystalline Materials ◽  
Substrate Interactions ◽  
Cell Substrate ◽  
Substrate Engineering ◽  
And Control

The aim of this study is to investigate the effect of atomic structure of polycrystalline materials on cell-substrate interactions. Samples are prepared from rods and sheets of Ti -6 Al -4 V substrates with predominately two distinct crystallographic orientations as well as nano-structured and annealed titanium fabricated through high-pressure torsion and heat treatment processes. The degree of preosteoblast attachment and rate of growth, which are regulated through the activity and interaction of proteins present in the extracellular matrix, are notably increased on the nano-structured titanium and substrate having predominant [Formula: see text] orientation. The improved cell activity is attributed to the nano-structured feature of these substrates consisting of ultra-fine crystals (< 50 nm) and specific atomic order of [Formula: see text] substrate which provide higher degree of surface wettability. These findings demonstrate the advantages of nano-structured titanium over the conventional and coated titanium implants, as both mechanical properties and cellular response are improved. Furthermore, crystal orientation of the substrates can influence cell responses and, therefore, substrate engineering can be used to improve and control cell-substrate interactions.

scholarly journals 136 EVIDENCE OF A DIRECT EFFECT OF P4 ON IVF-DERIVED BOVINE 8-CELL EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 219 ◽  
Author(s):  
C.E. Ferguson ◽  
T.R. Davidson ◽  
M.R.B. Mello ◽  
A.S. Lima ◽  
D.J. Kesler ◽  
...  
Keyword(s):  
Embryo Development ◽  
Direct Effect ◽  
Blastocyst Stage ◽  
Control Group ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Direct Role ◽  
Treatment Groups ◽  
And Control

There has been much debate over a direct role for progesterone (P4) in early bovine embryo development. While previous attempts to supplement bovine embryos in vitro with P4 produced results that vary and are often contradictory, this may be a response of administering P4 at inappropriate times. Therefore, the objective of these experiments was to determine if P4 could exert a direct effect on developing IVF-derived bovine embryos when administered at an appropriate time of embryo development. In Exp. I, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 168); (2) vehicle, CR1aa + ETOH (0.01%) (n = 170); and (3) P4, CR1aa + ETOH + P4 (20 ng/mL in 50-μL droplet) (n = 173). In Exp. II, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 160); (2) vehicle, CR1aa + DMSO (0.01%) (n = 180); and (3) P4, CR1aa + DMSO (0.01%) + P4 (20 ng/mL in 50-μL droplet) (n = 170). All embryos were evaluated on Days 6 to 9 post-insemination and rates calculated from 8-cell embryos. In Exp. I, ETOH tended to have a detrimental effect with significantly fewer (P < 0.05) embryos (53%) developing to the blastocyst stage on Day 7 compared with the control (62%) and P4 (71%) groups. At Day 7, significantly more embryos cultured in P4 (71%) developed to the blastocyst stage compared with the control group (62%). P4 treatment significantly increased the number of Grade 1 blastocysts (25%) on Day 7 compared with vehicle (15%) and control (17%) groups. At the end of culture, there were also significantly more Day 9 hatched blastocysts in the P4 group (33%) compared with vehicle (22%) and control (21%) groups. Supplementing P4 in the culture medium increased the rate of development, resulting in significantly more blastocysts (8%) on Day 6 and hatched blastocysts (21%) on Day 8 compared with vehicle (3% and 12%) and control (0% and 8%) groups, respectively. In Exp. II, there were no significant differences between treatment groups for Day 7 blastocysts (control 54%, DMSO 61%, P4 57%) and Day 9 hatched blastocysts (control 46%, DMSO 51%, P4 46%). However, there were significantly more Grade 1 blastocysts in the P4 group (22% and 36%) on Days 6 and 8 compared with vehicle (11% and 23%) and control (13% and 23%) groups, respectively. The lack of improvement in Day 7 blastocysts and Day 9 hatched blastocysts rates leads to further uncertainty in understanding the P4 vehicle interactions. In conclusion, the results of these two experiments indicate that P4 can exert a direct effect on the developing IVF-derived bovine embryo; however, due to P4 vehicle interactions; other inert vehicles need to be explored to further evaluate the direct effects of P4 on the developing bovine embryo.

Coenzyme A metabolism

1985 ◽  
Vol 248 (1) ◽  
pp. E1-E9 ◽  
Author(s):  
J. D. Robishaw ◽  
J. R. Neely
Keyword(s):  
Enzyme Activity ◽  
Cardiac Muscle ◽  
Heart Muscle ◽  
Coenzyme A ◽  
Free Carnitine ◽  
Pantothenate Kinase ◽  
Acetyl Coenzyme A ◽  
Synthetic Pathway ◽  
And Control ◽  
Inhibited Enzyme

The metabolism of coenzyme A and control of its synthesis are reviewed. Pantothenate kinase is an important rate-controlling enzyme in the synthetic pathway of all tissues studied and appears to catalyze the flux-generating reaction of the pathway in cardiac muscle. This enzyme is strongly inhibited by coenzyme A and all of its acyl esters. The cytosolic concentrations of coenzyme A and acetyl coenzyme A in both liver and heart are high enough to totally inhibit pantothenate kinase under all conditions. Free carnitine, but not acetyl carnitine, deinhibits the coenzyme A-inhibited enzyme. Carnitine alone does not increase enzyme activity. Thus changes in the acetyl carnitine-to-carnitine ratio that occur with nutritional states provides a mechanism for regulation of coenzyme A synthetic rates. Changes in the rate of coenzyme A synthesis in liver and heart occurs with fasting, refeeding, and diabetes and in heart muscle with hypertrophy. The pathway and regulation of coenzyme A degradation are not understood.

scholarly journals Coupled structural transitions enable highly cooperative regulation of human CTPS2 filaments

2019 ◽  
Author(s):  
Eric M. Lynch ◽  
Justin M. Kollman
Keyword(s):  
Enzyme Activity ◽  
Active Sites ◽  
Large Scale ◽  
Conformational State ◽  
Ctp Synthase ◽  
Widespread Phenomenon ◽  
And Control ◽  
Allosteric Regulators ◽  
Low Activity

Many enzymes assemble into defined oligomers, providing a mechanism for cooperatively regulating enzyme activity. Recent studies in tissues, cells, and in vitro have described a mode of regulation in which enzyme activity is modulated by polymerization into large-scale filaments1–5. Enzyme polymerization is often driven by binding to substrates, products, or allosteric regulators, and tunes enzyme activity by locking the enzyme in high or low activity states1–5. Here, we describe a unique, ultrasensitive form of polymerization-based regulation employed by human CTP synthase 2 (CTPS2). High-resolution cryoEM structures of active and inhibited CTPS2 filaments reveal the molecular basis of this regulation. Rather than selectively stabilizing a single conformational state, CTPS2 filaments dynamically switch between active and inactive filament forms in response to changes in substrate and product levels. Linking the conformational state of many CTPS2 subunits in a filament results in highly cooperative regulation, greatly exceeding the limits of cooperativity for the CTPS2 tetramer alone. The structures also reveal a link between conformational state and control of ammonia channeling between the enzyme’s two active sites. This filament-based mechanism of enhanced cooperativity demonstrates how the widespread phenomenon of enzyme polymerization can be adapted to achieve different regulatory outcomes.

scholarly journals Soil enzyme activity in the dry valley phytocenoses under the influence of Acer negundo L.

2021 ◽  
Vol 31 ◽  
pp. 00029
Author(s):  
Oksana Tsandekova
Keyword(s):  
Enzyme Activity ◽  
Hydrolytic Enzymes ◽  
Soil Condition ◽  
Invertase Activity ◽  
Horizontal Differentiation ◽  
Natural Ecosystems ◽  
Crown Density ◽  
Dry Valley ◽  
Medium Activity ◽  
And Control

The activity of hydrolytic enzymes in the soil of dry valley phytocenoses under the influence of ash-leaved maple was investigated. The research objects were selected taking into account the ranking of plantations by crown density. Soil samples were collected depending on the horizontal differentiation of communities in the undercrown and outer zones of phytogenic fields. An increase in the enzyme activity during the period of active tree growth among experimental and control samples was established. Among the enzymes, invertase demonstrated the highest activity, while protease and phosphatase were characterised by medium activity. An increased invertase activity was found in the trees with a high crown density as compared to the trees of other groups. The obtained data can be used as diagnostic indicators of soil condition for monitoring natural ecosystems.

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