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Biomedical subjects

W F Fong

Publications and source records attributed to W F Fong.

At least 37 records · Page 2Linked to original sources
PubMed1988

The induction of ornithine decarboxylase by L-asparagine and intracellular alkalinization.

The uptake of transport systems A and N amino acids, most noticeably L-asparagine, is essential for the induction of ornithine decarboxylase (L-ornithine carboxylase, EC 4.1.1.17) in cultured cells and we have proposed that the uptake-associated pH and ionic changes might constitute part of the cell activation signal (1). In the present study, it was shown that extracellular L-asparagine caused an immediate and transient increase in intracellular pH which was continuously monitored by the fluorescence probe BCECF (2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein). NH4Cl and NH4OH which caused intracellular alkalinization also caused ornithine decarboxylase activity to increase.

Ammonium Chloride↗
PubMed1987

Ornithine decarboxylase from Tetrahymena thermophila: purification, reversible modification and multiple forms.

T. thermophila ornithine decarboxylase was purified 1100-fold by an improved procedure to a specific activity of 2.31 x 10(4) nmol/hr/mg protein with a 28% yield. The purified enzyme was a monomer of Mr = 68,000 and had two isoelectric forms with pl's of 5.14 and 5.17 respectively. The Mr = 68,000 enzyme was reversibly modified by a cellular factor of approx. Mr = 10,000 to give rise to two additional forms with decreased DEAE-cellulose affinity.

Animals↗
PubMed1987

Asparagine transport and the induction of ornithine decarboxylase in H-35 rat hepatoma cells.

A number of amino acids, most noticeably asparagine, were capable of inducing ornithine decarboxylase in H-35 rat hepatoma cells. The effective amino acids were all neutral and were substrates of the Na+-dependent transport systems A, ASC and N. Transport inhibitor studies indicated that there was an excellent correlation between the level of enzyme activity induced and the initial rate of asparagine transport into the cells by the A and the N systems. It is proposed that the activation of the Na+-dependent, pH-sensitive amino acid transport systems and the subsequent intracellular pH and ionic perturbation constitute part of the initiation signals for cell activation.

Amino Acids↗
PubMed1985

The effect of transport system A and N amino acids and of nerve and epidermal growth factors on the induction of ornithine decarboxylase activity.

The induction of ornithine decarboxylase (EC 4.1.1.17) (ODC) by amino acids and by the peptide hormones nerve growth factor (NGF) and epidermal growth factor (EGF) in salts-glucose media has been studied. Only those neutral amino acids taken into the cell via one of the Na+ dependent transport systems stimulate ODC activity. Asparagine and the nonmetabolizable alpha-amino-isobutyric acid (AIB) were used as representatives of this class of inducing amino acids, and their intracellular concentrations were related to the levels of ODC induced. A threshold intracellular concentration of asparagine or AIB has to be attained before ODC can be induced. Further slight increases in intracellular concentrations of asparagine or AIB produce disproportionately large increases of ODC, resulting in a sigmoidal curve of ODC induction. These results, and the fact that the decrease in ODC levels caused by valine is associated with a concurrent decrease in the intracellular level of the inducing amino acid, suggest that the intracellular amino acid level is causally related to the induction of ornithine decarboxylase. Glutamic acid, EGF, and NGF do not induce ODC except in the presence of an inducing amino acid. They act synergistically with the inducing amino acid and produce higher ODC levels at the same intracellular concentration of the inducing amino acid.

Amino Acids↗
PubMed1985

Regulation of polyamine biosynthesis by antizyme and some recent developments relating the induction of polyamine biosynthesis to cell growth. Review.

This review considers the role of antizyme, of amino acids and of protein synthesis in the regulation of polyamine biosynthesis. The ornithine decarboxylase of eukaryotic cells and of Escherichia coli can be non-competitively inhibited by proteins, termed antizymes, which are induced by di- and poly- amines. Some antizymes have been purified to homogeneity and have been shown to be structurally unique to the cell of origin. Yet, the E. coli antizyme and the rat liver antizyme cross react and inhibit each other's biosynthetic decarboxylases. These results indicate that aspects of the control of polyamine biosynthesis have been highly conserved throughout evolution. Evidence for the physiological role of the antizyme in mammalian cells rests upon its identification in normal uninduced cells, upon the inverse relationship that exists between antizyme and ornithine decarboxylase as well as upon the existence of the complex of ornithine decarboxylase and antizyme in vivo. Furthermore, the antizyme has been shown to be highly specific; its Keq for ornithine decarboxylase is 1.4 X 10(11) M-1. In addition, mammalian cells contain an anti-antizyme, a protein that specifically binds to the antizyme of an ornithine decarboxylase-antizyme complex and liberates free ornithine decarboxylase from the complex. In E. coli, in which polyamine biosynthesis is mediated both by ornithine decarboxylase and by arginine decarboxylase, three proteins (one acidic and two basic) have been purified, each of which inhibits both these enzymes. They do not inhibit the biodegradative ornithine and arginine decarboxylases nor lysine decarboxylase. The two basic inhibitors have been shown to correspond to the ribosomal proteins S20/L26 and L34, respectively. The relationship of the acidic antizyme to other known E. coli proteins remains to be determined. In mammalian cells, ornithine decarboxylase can be induced by a broad spectrum of compounds. These range from hormones and growth factors to natural amino acids such as asparagine and to non-metabolizable amino acid analogues such as alpha-amino-isobutyric acid. The amino acids that induce ornithine decarboxylase as well as those that promote polyamine uptake utilize the sodium dependent A and N transport systems. Consequently, they act in concert and increase intracellular polyamine levels by both mechanisms. The induction of ornithine decarboxylase by growth factors, such as NGF, EGF, and PDGF as well as by insulin requires the presence of these same amino acids and does not occur in their absence. However, the inducing amino acid need not be incorporated into protein nor covalently modified.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed1985

Changes in ornithine decarboxylase activity and polyamine levels during the growth of Tetrahymena thermophila cultures.

Ornithine decarboxylase activity and polyamine levels were determined at various growth phases of Tetrahymena thermophila cultures. Enzyme activity and intracellular polyamines increased in exponentially growing cells and peaked just before the stationary phase. Putrescine was the predominant polyamine and spermidine and spermine concentrations were low throughout. The increase in putrescine level can be totally accounted for by the enzyme activity detected, provided that there is an ample supply of the precursor, L-ornithine.

Animals↗
PubMed1985

Effects of spermine on transferrin and iron uptake by reticulocytes: I. Action on the endocytic uptake of transferrin.

Iron uptake by rabbit reticulocytes was inhibited by spermine in a concentration-dependent manner. Examination of the single-cycle endocytosis of 125I-transferrin showed that a graded reduction in the rate of exocytosis of transferrin was related to increasing extracellular spermine concentrations. This reduction could affect the recycling of transferrin receptors and resulted in the loss of membrane binding sites in spermine-treated cells. As large vacuoles were observed in cells treated with spermine, the endotubular function of these cells was probably affected. Spermine also enhanced the binding affinity of transferrin to membrane receptors. The mechanism for this enhancement was not clear.

Animals↗
PubMed1985

Effects of spermine on transferrin and iron uptake by reticulocytes: II. Changes in intravesicular pH.

An increase in extracellular spermine concentration brought about a progressive rise in intralysosomal pH in rabbit reticulocytes. Since intracellular release of iron from transferrin is believed to involve the protonation of the iron-transferrin complex, the rise in intralysomal pH could account for the inhibitory effect of spermine on iron uptake. The inhibition could be reversed if spermine was removed by washing. As a result of spermine treatment, more acid-labile N-terminal monoferric transferrin and less apotransferrin were released from the cell. These results are consistant with the protonation theory of iron release.

Animals↗
PubMed1985

Limited blood-brain barrier transport of polyamines.

Transport of polyamines across the blood-brain barrier of adult rats was examined by measuring the amount of radioactivity that reached the forebrain 5 s after a "bolus" intracarotid injection. The values were expressed by the brain uptake index (BUI), which is the percentage of material transported in relation to freely diffusible water in a single passage through the brain. Transport was restricted as indicated by the respective BUI values, presented as means +/- SD (number of animals): putrescine, 5.3 +/- 0.8 (11); spermidine, 6.1 +/- 1.3 (7); and spermine, 5.8 +/- 0.5 (4). A kinetic study of the transport of [14C]putrescine showed that transport due to passive diffusion accounted for the majority of the observed influx (66% at 1 mM putrescine). However, a small saturable component exists with a Km value of 4-5 mM and a Vmax of 30 nmol X min-1 X g-1. This Km value is considerably higher than the circulating levels of the polyamine in the normal mature animal, and thus is unlikely to be of physiological significance. Competition studies indicated that putrescine does not interact with carriers for adenosine, arginine, choline, or leucine.

Adenosine↗
PubMed1984

Putrescine biosynthesis in Tetrahymena thermophila.

The putrescine-biosynthesis pathway in Tetrahymena thermophila was delineated by studying crude extracts prepared from exponentially growing cultures. A pyridoxal phosphate-stimulated ornithine decarboxylase activity competitively inhibited by putrescine was detected. CO2 was also liberated from L-arginine, but analyses by t.l.c. and enzyme studies suggested that the activity was not due to arginine decarboxylase, nor could enzyme activities converting agmatine into putrescine be detected. We conclude that the decarboxylation of L-ornithine is probably the only major route for putrescine biosynthesis in this organism during exponential growth.

Agmatine↗
PubMed1984

Effects of polyamines on the uptake of neurotransmitters by rat brain synaptosomes.

The influence of putrescine, spermidine, spermine, and some aliphatic alpha, w-diamines on the uptake spermine, and some aliphatic alpha, w-diamines on the uptake of neurotransmitters by rat forebrain synaptosomes was investigated. Choline uptake was most effectively inhibited by spermine (IC50 = 0.22 mM), less so by spermidine (IC50 = 4.0 mM), but not by putrescine (IC50 greater than 100 mM). At 10 mM, 1,3-diaminopropane, cadaverine, and 1,8-diaminooctane all inhibited choline uptake by 50% or more. Spermine and spermidine inhibited the uptake of dopamine with IC50 values of 2.7 and 2.2 mM, respectively. Putrescine was only slightly inhibitory (IC50 = 17.3 mM) and the other diamines were inactive. The uptake of gamma-aminobutyrate (GABA) was only slightly inhibited (15-40%) by the polyamines at 10 mM. With the exception of inhibition of glycine uptake by 1,8-diaminooctane (60%) and of glutamate uptake by cadaverine (35%) none of the polyamines, tested at 10 mM, affected the uptake of adenosine, glutamate, and glycine significantly. A possible modulatory role for polyamines in synaptic transmission through interaction by negatively charged groups of the synaptic membrane with the polycationic compounds is discussed.

Animals↗
PubMed1976

The appearance of an ornithine decarboxylase inhibitory protein upon the addition of putrescine to cell cultures.

Quiescent, contact inhibited H-35 rat hepatoma cell cultures maintained in minimal essential medium contain a very low level of ornithine decarboxylase activity. However, 2 h after the addition of 10% fetal calf serum to the culture medium, the enzyme activity increases by approx. 100-fold. This increase can be completely inhibited by the simultaneous addition of 10(-2) M putrescine. The presence of putrescine elicits the appearance of an intracellular inhibitor of ornithine decarboxylase. This inhibitor of ornithine decarboxylase has a molecular weight of 26500, is sensitive to the action of chymotrypsin and is noncompetitive with respect to ornithine. The intracellular appearance of this inhibitor is sensitive to cycloheximide but is only partially inhibited by actinomycin D.

Carboxy-Lyases↗
PubMed1976

Studies on the mode of action of ecdysterone in adult female Aedes aegypti.

Injection of ecdysterone into non-blood fed adult female Aedes aegypti results in a marked stimulation of aromatic-L-amino-acid decarboxylase (formerly DOPA-decarboxylase) activity (Schlaeger and Fuchs, 1974a). When the hormone and alpha-amanitin are injected either simultaneously or if the toxin is administered first no inhibition of subsequent enzymatic activity is observed and in fact substantial enhancement occurs. Cordycepin injection along with ecdysterone gives results similar to alpha-amanitin. The inhibitors by themselves elicit a very small increase in DOPA decarboxylase activity compared to saline-injected controls. Conversely, actinomycin D causes severe depression of ecdysterone-mediated DOPA decarboxylase activity as dose cycloheximide and puromycin. We interpret our data to mean that the mRNA for DOPA decarboxylase is already present prior to exposure to ecdysterone. We postulate that the function of the hormone would be to modulate translation of specific pre-formed mRNA's by an unknown mechanism or to induce transcription of specific tRNAs necessary for the initation of translation of selecting existing messengers.

Aedes↗