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Biomedical subjects

Keith F Tipton

Publications and source records attributed to Keith F Tipton.

12 recordsLinked to original sources
PubMed2006

The therapeutic potential of monoamine oxidase inhibitors.

Monoamine oxidase inhibitors were among the first antidepressants to be discovered and have long been used as such. It now seems that many of these agents might have therapeutic value in several common neurodegenerative conditions, independently of their inhibition of monoamine oxidase activity. However, many claims and some counter-claims have been made about the physiological importance of these enzymes and the potential of their inhibitors. We evaluate these arguments in the light of what we know, and still have to learn, of the structure, function and genetics of the monoamine oxidases and the disparate actions of their inhibitors.

Animals↗
PubMed2005

Structure-activity studies with high-affinity inhibitors of pyroglutamyl-peptidase II.

Inhibitors of PPII (pyroglutamyl-peptidase II) (EC 3.4.19.6) have potential applications as investigative and therapeutic agents. The rational design of inhibitors is hindered, however, by the lack of an experimental structure for PPII. Previous studies have demonstrated that replacement of histidine in TRH (thyrotropin-releasing hormone) with asparagine produces a competitive PPII inhibitor (Ki 17.5 microM). To gain further insight into which functional groups are significant for inhibitory activity, we investigated the effects on inhibition of structural modifications to Glp-Asn-ProNH2 (pyroglutamyl-asparaginyl-prolineamide). Synthesis and kinetic analysis of a diverse series of carboxamide and C-terminally extended Glp-Asn-ProNH2 analogues were undertaken. Extensive quantitative structure-activity relationships were generated, which indicated that key functionalities in the basic molecular structure of the inhibitors combine in a unique way to cause PPII inhibition. Data from kinetic and molecular modelling studies suggest that hydrogen bonding between the asparagine side chain and PPII may provide a basis for the inhibitory properties of the asparagine-containing peptides. Prolineamide appeared to be important for interaction with the S2' subsite, but some modifications were tolerated. Extension of Glp-Asn-ProNH2 with hydrophobic amino acids at the C-terminus led to a novel set of PPII inhibitors active in vitro at nanomolar concentrations. Such inhibitors were shown to enhance recovery of TRH released from rat brain slices. Glp-Asn-Pro-Tyr-Trp-Trp-7-amido-4-methylcoumarin displayed a Ki of 1 nM, making it the most potent competitive PPII inhibitor described to date. PPII inhibitors with this level of potency should find application in exploring the biological functions of TRH and PPII, and potentially provide a basis for development of novel therapeutics.

Amino Acid Sequence↗
PubMed2005

Characterization of cinnamyl alcohol dehydrogenase of Helicobacter pylori. An aldehyde dismutating enzyme.

Cinnamyl alcohol dehydrogenases (CAD; 1.1.1.195) catalyse the reversible conversion of p-hydroxycinnamaldehydes to their corresponding alcohols, leading to the biosynthesis of lignin in plants. Outside of plants their role is less defined. The gene for cinnamyl alcohol dehydrogenase from Helicobacter pylori (HpCAD) was cloned in Escherichia coli and the recombinant enzyme characterized for substrate specificity. The enzyme is a monomer of 42.5 kDa found predominantly in the cytosol of the bacterium. It is specific for NADP(H) as cofactor and has a broad substrate specificity for alcohol and aldehyde substrates. Its substrate specificity is similar to the well-characterized plant enzymes. High substrate inhibition was observed and a mechanism of competitive inhibition proposed. The enzyme was found to be capable of catalysing the dismutation of benzaldehyde to benzyl alcohol and benzoic acid. This dismutation reaction has not been shown previously for this class of alcohol dehydrogenase and provides the bacterium with a means of reducing aldehyde concentration within the cell.

Alcohol Oxidoreductases↗
PubMed2004

Electrochemical investigation into the redox activity of Fe(II)/Fe(III) in the presence of nicotine and possible relations to neurodegenerative diseases.

The biological relevance of Fe(II)/Fe(III) is becoming evermore apparent, especially in relation to its potential role in the progression of neurodegenerative diseases such as Parkinson's and Alzheimer's disease. The reported relationship between smoking and a reduced incidence of neurodegenerative disorders prompted this work. In order to investigate whether nicotine can interact with iron, we have studied the electrochemical behaviour of a Fe(II)/Fe(III) redox couple in the presence of nicotine. Solubility issues and lack of available nonreacting salts of nicotine necessitated studies being conducted at low pH values. Cyclic voltammetry experiments revealed a definite alteration in the electrochemical behaviour of the Fe(II)/Fe(III) redox couple suggesting the capability of nicotine to complex with free iron and, hence, reduce its reactivity. This is evident from a slower rate of heterogeneous electron transfer, ks, and a shift from reversible to quasi-reversible behaviour, as characterised from the diffusion coefficient (D), the full width half maximum (FWHM), DeltaEp and Ef. Additional complexation titrations, pH ranging from 1 to 7, confirm a weak complexation reaction occurring between Fe(III) and nicotine.

Electrochemistry↗
PubMed2004

Taurine prevents streptozotocin impairment of hormone-stimulated glucose uptake in rat adipocytes.

Streptozotocin-treated rats were used as models of type 1 diabetes to study the effects of dietary taurine on insulin- and adrenergic-stimulated 2-deoxyglucose uptake by isolated adipocytes. In addition to the well-established impairment of basal and insulin-stimulated 2-deoxyglucose uptakes in adipocytes prepared from streptozotocin-diabetic rats, the alpha-(phenylephrine) and beta-(isoproterenol) adrenergic stimulations of glucose uptake were also abolished. The insulin stimulation of glucose uptake in adipocytes was selectively abolished by the phosphatidylinositol 3-kinase inhibitor wortmannin, whereas that by the adrenergic agonists, phenylephrine and isoproterenol, was inhibited by prazosin and propranolol, respectively. Dietary taurine, 4 weeks before and 4 weeks after streptozotocin administration, prevented the loss of both insulin and adrenergic agonist stimulations of 2-deoxyglucose uptake, without affecting hyperglycaemia. Because insulin and adrenergic activations of glucose transport by adipocytes are coupled to different signalling pathways, it is unlikely that these effects of taurine are related to these disparate postreceptor mechanisms.

Adipocytes↗
PubMed2004

IntEnz, the integrated relational enzyme database.

IntEnz is the name for the Integrated relational Enzyme database and is the official version of the Enzyme Nomenclature. The Enzyme Nomenclature comprises recommendations of the Nomenclature Committee of the International Union of Bio chemistry and Molecular Biology (NC-IUBMB) on the nomenclature and classification of enzyme-catalysed reactions. IntEnz is supported by NC-IUBMB and contains enzyme data curated and approved by this committee. The database IntEnz is available at http://www.ebi.ac.uk/intenz.

Animals↗
PubMed2004

Semicarbazide-sensitive amine oxidases: enzymes with quite a lot to do.

The semicarbazide-sensitive amine oxidases (SSAO) (EC 1.4.3.6) were believed to be detoxifying enzymes, primarily involved in the oxidative deamination of endogenous amines, such as methylamine and aminoacetone, together with some xenobiotic amines. However, it appears that the reaction products may have important signalling functions in the regulation of cell development and glucose homeostasis. Furthermore, enzyme, from some sources, behaves as a cellular adhesion protein under inflammatory and it may also be involved in lipid transport. This review considers what is known about the activities and potential functions of this hardworking protein.

Amine Oxidase (Copper-Containing)↗
PubMed2004

Monoamine oxidases: certainties and uncertainties.

A great deal has been learned about the behaviour of monoamine oxidase in the 75 years since it was first discovered, but there is still a great deal left to understand. This review concentrates on the dynamic aspects of our knowledge of the interactions of MAO with substrates and inhibitors and how it may collaborate with other enzymes, with particular emphasis on aspects that remain to be clarified.

Animals↗
PubMed2003

Semicarbazide-sensitive amine oxidases in pig dental pulp.

The behaviour of semicarbazide-sensitive amine oxidase (SSAO; E.C. 1.4.3.6) in dental pulp has been studied, with particular reference to the metabolism of 5-hydroxytryptamine (5-HT; serotonin). Kinetic studies using radioactively labelled substrates have confirmed benzylamine, 2-phenylethylamine (PEA) and 5-HT to be substrates for microsomal SSAO from porcine dental pulp. Kinetic substrate-competition studies indicated the presence of two forms of SSAO in dental pulp; one that oxidises benzylamine and PEA but not 5-HT and a second that oxidises 5-HT and PEA but not benzylamine. These two forms also differ in their thermostabilities at 60 and 70 degrees C, although this thermal inactivation is partly reversible.

Amine Oxidase (Copper-Containing)↗
PubMed2003

The inhibition of semicarbazide-sensitive amine oxidase by aminohexoses.

Semicarbazide-sensitive amine oxidase (EC 1.4.3.6; amine:oxygen oxidoreductase (deaminating) (copper-containing); SSAO) is a multifunctional protein. It acts under inflammatory conditions as a vascular-adhesion protein (VAP-1), mediating the adhesion of lymphocytes to vascular endothelial cells. The relationships, if any, between this adhesion function and the enzymatic functions (amine-substrate specificity and catalysis) of SSAO have not yet been defined. Since cell surface amino sugars and their derivatives are known to be involved in cell-to-cell recognition, we have investigated their possible effects on the enzyme activity of SSAO. The aminohexoses galactosamine, glucosamine and mannosamine were not oxidatively deaminated by SSAO. However, their presence during the assay of benzylamine oxidation resulted in a time-dependent inhibition. This inhibition was shown to follow saturation kinetics with respect to hexosamine concentration. Although time-dependent, the inhibition of SSAO activity was found to be reversible by dilution. In contrast, there is no such inhibition when the N-acetylamino sugar derivatives or the parent sugars (galactose, glucose and mannose) replaced the amino sugars in the reaction mixture. These results suggest that the interactions between SSAO and aminohexoses are specific and, therefore, that the cell-adhesion functions and amine-recognition functions of VAP-1/SSAO may be interlinked.

Amine Oxidase (Copper-Containing)↗
PubMed2002

Interactions of the neurotoxin 6-hydroxydopamine with glyceraldehyde-3-phosphate dehydrogenase.

The neurotoxin 6-hydroxydopamine (6-OHDA) oxidised sulphydryl groups in glyceraldenyde-3-phosphate dehydrogenase (GAPDH) with loss of the dehydrogenase activity of the enzyme. 5-Hydroxydopamine behaved similarly at higher concentrations. This oxidation was accompanied by a transient rise in the acyl phosphatase (non-phosphorylating) activity of GAPDH. Treatment with arsenite resulted in loss of the acyl phosphatase activity and restoration of the dehydrogenase activity, consistent with the process involving sulphydryl-group oxidation to the corresponding sulphenate. Prolonged incubation with 6-OHDA resulted in a loss of both enzyme activities. Arsenite was unable to reverse this inhibition, indicating further oxidation, perhaps to sulphinate and sulphonate, to have occurred. RNA, but not DNA, was a partial inhibitor of both the dehydrogenase and acyl phosphatase activities of GAPDH, whereas DNA (both single- and double-stranded) was ineffective. Both single-stranded DNA and RNA inhibited the esterase activity of GAPDH, an activity that requires the absence of NAD+, in a process that was relieved at higher polynucleotide concentrations. Except at very high concentrations (1 mM), treatment of GAPDH 6-OHDA was ineffective at abolishing its to single-stranded DNA. Since GAPDH is an essential enzyme in glycolysis and also plays a role in apoptotic cell death, these results suggest that the effects on this enzyme may contribute to the neurotoxicity of 6-OHDA.

Adrenergic Agents↗