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A T Brown

Publications and source records attributed to A T Brown.

50 records · Page 3Linked to original sources
PubMed1973

Identification, preliminary characterization, and evidence for regulation of invertase in Streptococcus mutans.

Sucrose dissimilation was studied in five strains of Streptococcus mutans. Glucose-adapted strain SL-1 makes acid more slowly from sucrose than from glucose; glucose-adapted strain SL-1 gives diauxie growth kinetics in broth containing limiting amounts of both glucose and sucrose. Thus, at least part of the sucrose dissimilative system appears inducible. Sucrase activity was identified in the 37,000 x g soluble cell fraction of five strains. Its intracellular location implies the presence of sucrose permease. The specific activity of the sucrase is higher in sucrose-adapted cells than in cells adapted to glucose or other sugars, further suggesting its inducibility. The enzyme from strain SL-1 was partially purified by diethylaminoethyl-cellulose chromatography and shown to be a single molecule with a molecular weight of about 48,000. The partially purified enzyme is specific for sucrose and produces equimolar glucose and fructose. Since it degrades raffinose, but not melezitose or other alpha-glucosides, it is an invertase. The invertase has a relatively high K(m) for its substrate and a pH optimum of 5.5 to 6.2. It is activated by inorganic orthophosphate (P(i)), P(i) functioning as a positive effector. Arsenate can substitute for phosphate. Neither the crude cell-free extract nor the partially purified enzyme preparations has detectable sucrose phosphorylase activity. A possible potent role of the invertase in the regulation of sucrose carbon flow in S. mutans is discussed.

Acids↗
PubMed1972

Induction and regulation of a nicotinamide adenine dinucleotide-specific 6-phosphogluconate dehydrogenase in Streptococcus faecalis.

Streptococcus faecalis grown with glucose as the primary energy source contains a single, nicotinamide adenine dinucleotide phosphate (NADP)-specific 6-phosphogluconate dehydrogenase. Extracts of gluconate-adapted cells, however, exhibited 6-phosphogluconate dehydrogenase activity with either NADP or nicotinamide adenine dinucleotide (NAD). This was shown to be due to the presence of separate enzymes in gluconate-adapted cells. Although both enzymes catalyzed the oxidative decarboxylation of 6-phosphogluconate, they differed from one another with respect to their coenzyme specificity, molecular weight, pH optimum, K(m) values for substrate and coenzyme, and electrophoretic mobility in starch gels. The two enzymes also differed in their response to certain effector ligands. The NADP-linked enzyme was specifically inhibited by fructose-1,6-diphosphate, but was insensitive to adenosine triphosphate (ATP) and certain other nucleotides. The NAD-specific enzyme, in contrast, was insensitive to fructose-1,6-diphosphate, but was inhibited by ATP. The available data suggest that the NAD enzyme is involved primarily in the catabolism of gluconate, whereas the NADP enzyme appears to function in the production of reducing equivalents (NADPH) for use in various reductive biosynthetic reactions.

Adenosine Triphosphate↗
PubMed1972

Fructose-1,6-diphosphate-dependent lactate dehydrogenase from a cariogenic streptococcus: purification and regulatory properties.

The lactate dehydrogenase (LDH) from Streptococcus mutans NCTC 10449 is under stringent metabolic control. The partially purified enzyme was specifically activated by high concentrations of fructose-1,6-diphosphate (FDP) and was inhibited by adenosine triphosphate. There appeared to be at least two binding sites for the activator which interacted in a cooperative manner. The interaction between the FDP sites was independent of the pH of the assay system, although the relative affinity of the enzyme for the activator was influenced by pH. There also appeared to be at least two pyruvate binding sites on the S. mutans LDH with some cooperative interaction between them, and the interaction between these sites was also independent of the hydrogen ion concentration. Two pyruvate analogues had different effects on the interaction of pyruvate with the LDH. One of the analogues, alpha-ketobutyrate, stimulated enzyme activity at limiting pyruvate concentrations, but had no significant effect at saturating concentrations of the substrate. The net effect of alpha-ketobutyrate was to shift the pyruvate saturation curve from sigmoidal to hyperbolic and to decrease the Hill coefficient from about 2.0 to 1.0. The other pyruvate analogue, oxamate, inhibited enzyme activity at all pyruvate concentrations but had no effect on the sigmoidal nature of the pyruvate saturation curve or on the apparent kinetic order of the reaction with respect to substrate. These results suggested that there may be two types of pyruvate binding sites on the LDH from S. mutans. Other kinetic properties of the S. mutans NCTC 10449 enzyme were studied and compared with those exhibited by the LDH from several other strains of the organism.

Adenosine Triphosphate↗
PubMed1971

Mechanism for regulating the distribution of glucose carbon between the Embden-Meyerhof and hexose-monophosphate pathways in Streptococcus faecalis.

Glucose-adapted Streptococcus faecalis produced little if any (14)CO(2) from glucose-1-(14)C, although high levels of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) were detected in cell-free extracts. Metabolism of glucose through the oxidative portion of the hexose-monophosphate pathway was shown to be regulated in this organism by the specific inhibitory interaction of the Embden-Meyerhof intermediate, fructose-1, 6-diphosphate (FDP), with 6-phosphogluconate dehydrogenase. Glucose-6-phosphate dehydrogenase activity was unaffected by FDP. The S. faecalis 6-phosphogluconate dehydrogenase was partially purified from crude extracts by standard fractionation procedures and certain kinetic parameters of the FDP-mediated inhibition were investigated. The negative effector was shown to cause a decrease in V(max) and an increase in the apparent K(m) for both 6-phosphogluconate and nicotinamide adenine dinucleotide phosphate (NADP). These effects were apparently a consequence of the ligand interacting with the enzyme at a site distinct from either the substrate or the coenzyme sites. Among the evidence supporting this was the fact that beta-mercaptoethanol blocked completely FDP inhibition, but had no effect on catalytic activity. The possibility that the regulation of 6-phosphogluconate dehydrogenase activity by FDP might be of some general significance was suggested by the observation that this enzyme from several other sources was also sensitive to FDP.

Animals↗
PubMed1970

Regulation of enzymes involved in the conversion of tryptophan to nicotinamide adenine dinucleotide in a colorless strain of Xanthomonas pruni.

A colorless strain of Xanthomonas pruni was isolated which is capable of converting tryptophan to nicotinamide adenine dinucleotide (NAD). The enzymes responsible for the conversion of tryptophan to quinolinic acid were shown to be present. Nicotinic acid-requiring mutants were isolated, and it was found that the growth of these mutants can be supported by various intermediates on the pathway from tryptophan to NAD. The first three enzymes on this pathway are induced coordinately by l-tryptophan. Gratuitous inducers of these enzymes include d-tryptophan, alpha-methyl-dl-tryptophan, and 4-methyl-dl-tryptophan; formyl-l-kynurenine and l-kynurenine were not effective as inducers. These data suggest that at least the first three enzymes in the pathway from tryptophan to NAD are under common regulatory control.

Amidohydrolases↗
PubMed1970

Regulation of tryptophan pyrrolase activity in Xanthomonas pruni.

Tryptophan pyrrolase was studied in partially purified extracts of Xanthomonas pruni. The dialyzed enzyme required both heme and ascorbate for maximal activity. Other reducing agents were able to substitute for ascorbate. Protoporphyrin competed with heme for the enzyme, suggesting that the native enzyme is a hemoprotein. The enzyme exhibited sigmoid saturation kinetics. Reduced nicotinamide adenine dinucleotide (NADH), reduced nicotinamide adenine dinucleotide phosphate (NADPH), nicotinic acid mononucleotide, and anthranilic acid enhanced the sigmoid kinetics and presumably bound to allosteric sites on the enzyme. The sigmoid kinetics were diminished in the presence of alpha-methyltryptophan. NAD, NADP, nicotinic acid, nicotinamide, nicotinamide mononucleotide, and several other related compounds were without effect on the activity of the enzyme. These data indicate that the activity of the enzyme is under feedback regulation by the ultimate end products of the pathway leading to NAD biosynthesis, as well as by certain intermediates of this pathway.

Ascorbic Acid↗
PubMedDate not supplied

Increased intimal hyperplasia after carotid endarterectomy in spontaneously hypertensive rats.

This study evaluated the effects of hypertension on postoperative intimal hyperplasia using a rat carotid endarterectomy (CEA) model with spontaneously hypertensive rats (SHR) and normotensive Sprague-Dawley rats (SD). SHR and SD rats underwent left carotid exposure and CEA via an arteriotomy, scoring and removal of the intima, followed by arteriotomy closure. The rats were then sacrificed two weeks postoperatively. The left carotid artery was harvested and underwent elastin and double immunohistochemical staining. The percent of lumenal stenosis was calculated using morphometric measurements, and stained cells within the intimal hyperplasia were counted. The means and standard deviation of the means were calculated, and the two groups were compared using a 2-sample t test. The systolic blood pressure was 228 +/-35 mm Hg in the SHR group and 108 +/-8 mm Hg in the SD group (p<0.00001). The percent of lumenal stenosis was 82.6 +/-17.1% in the SHR group and 21.2 +/-13.7% in the SD rats (p value <0.0001). The percentage of cells staining for a-SM actin was equal in the SHR group and the SD group (> 91%) but the percentage of these cells staining for BrdU was 38.2 +/-8.4% in the SHR group and 10.7 +/-5.8% in the SD group (p< 0.00001). Hypertension in the SHR rats was associated with an increased lumenal stenosis due to increased intimal hyperplasia. The increased intimal hyperplasia was due to enhanced and accelerated replication of smooth muscle cells. Hypertension may be associated with increased restenosis rates after CEA.

Actins↗