Hydrolysis of flavin-adenine dinucleotide by rat liver lysosomes

Ragab, Mohab; Brightwell, R.; Tappel, Al L.

doi:10.1016/0003-9861(68)90117-3

Cited by 23 publications

(9 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…mg protein-' was found. Interestingly, no increase in fluorescence was found when 1 mM AMP, an inhibitor of certain FAD pyrophosphatase (Ragab et al, 1968;Shin and Mego, 1988), was added together with FAD ( Fig. 1 A).…”

Section: Resultsmentioning

confidence: 96%

“…1 (seven experiments). The lysosomal contamination in purified RLM was checked by measuring the activities of the acid FAD pyrophosphatase and acid phosphatase, lysosomal enzymes associated with soluble fraction and both soluble and membrane fractions, respectively (Ragab et al, 1968 ;Shibko and Tappel, 1963). Acid FAD pyrophosphatase activity could not be detected in purified RLM, while the acid phosphatase specific activity was found to decrease by 90 2 7 % (five experiments) compared to that found in non-purified mitochondria.…”

Section: Methodsmentioning

confidence: 96%

“…Consistently, FAD assembly with imported apoenzymes was also reported (Nagao and Tanaka, 1992). On the other hand, both FAD and FMN are catabolised to riboflavin outside mitochondria (Ragab et al, 1968;McCormick, 1989;Kim et al, 1993). Whether RLM can metabolise FAD and FMN is at present unknown; however, in the light of the failure of FAD to enter mitochondria, the possibility must be taken in consideration that FAD and FMN catabolism occurs in mitochondria, maybe by the route of mitochondrial flavoprotein degradation.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Flavin Adenine Dinucleotide and Flavin Mononucleotide Metabolism in Rat Liver

Barile¹,

Brizio²,

Virgilio³

et al. 1997

European Journal of Biochemistry

View full text Add to dashboard Cite

In order to gain some insight into mitochondrial flavin biochemistry, rat liver mitochondria essentially free of lysosomal and microsomal contamination were prepared and their capability to metabolise externally added and endogenous FAD and FMN tested both spectroscopically and via HPLC.The existence of two novel mitochondrial enzymes, namely FAD pyrophosphatase (EC 3.6.1.18) and FMN phosphohydrolase (EC 3.1.3.2), which catalyse FAD+FMN and FMN-riboflavin conversion, respectively, is shown. They differ from each other and from extramitochondrial enzymes, as judged by their pH profile and inhibitor sensitivity, and can be separated in a partial FAD pyrophosphatase purification.Digitonin titration and subfractionation experiments show that FAD pyrophosphatase is located in the outer mitochondrial membrane and FMN phosphohydrolase in the intermembrane space. Since these enzymes can metabolise endogenous FAD and FMN, which are made available by using both Triton X-100 and the effector oxaloacetate, a proposal is made that FAD pyrophosphatase and FMN phosphohydrolase play a major role in mitochondrial flavoprotein turnover.

show abstract

Section: Resultsmentioning

confidence: 96%

Section: Methodsmentioning

confidence: 96%

mentioning

confidence: 99%

See 1 more Smart Citation

Flavin Adenine Dinucleotide and Flavin Mononucleotide Metabolism in Rat Liver

Barile¹,

Brizio²,

Virgilio³

et al. 1997

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…Nevertheless, the strong inhibition by AMP allows us to postulate that the dinucleotide was broken at the level of pyrophosphate bond, as already demonstrated for certain animal and plants FAD pyrophosphatases (FADppase, EC 3.6.1.18) [54, 55]. …”

Section: Resultsmentioning

confidence: 91%

A Regulatory Role of NAD Redox Status on Flavin Cofactor Homeostasis inS. cerevisiaeMitochondria

Locato

Barile

2013

Oxidative Medicine and Cellular Longevity

View full text Add to dashboard Cite

Flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NAD) are two redox cofactors of pivotal importance for mitochondrial functionality and cellular redox balance. Despite their relevance, the mechanism by which intramitochondrial NAD(H) and FAD levels are maintained remains quite unclear in Saccharomyces cerevisiae. We investigated here the ability of isolated mitochondria to degrade externally added FAD and NAD (in both its reduced and oxidized forms). A set of kinetic experiments demonstrated that mitochondrial FAD and NAD(H) destroying enzymes are different from each other and from the already characterized NUDIX hydrolases. We studied here, in some detail, FAD pyrophosphatase (EC 3.6.1.18), which is inhibited by NAD+ and NADH according to a noncompetitive inhibition, with Ki values that differ from each other by an order of magnitude. These findings, together with the ability of mitochondrial FAD pyrophosphatase to metabolize endogenous FAD, presumably deriving from mitochondrial holoflavoproteins destined to degradation, allow for proposing a novel possible role of mitochondrial NAD redox status in regulating FAD homeostasis and/or flavoprotein degradation in S. cerevisiae.

show abstract

“…The plasma enzyme, which hydrolyzes FAD to FMN, was assayed by the method described by Ragab et al (1968) unless otherwise indicated. To one volume of citrated plasma, three volumes of normal saline were added.…”

Section: Methodsmentioning

confidence: 99%

Further studies on the effects of flavine adenine dinucleotide on the platelet functions.

Shimada

Takahashi

Watanabe

et al. 1983

Tohoku J. Exp. Med.

View full text Add to dashboard Cite

Flavine adenine dinucleotide (FAD) may inhibit not only the aggregation but also ATP release of platelets in vitro. FAD did not inhibit the synthesis of thromboxane from 1-14C-arachidonic acid by platelets. FAD was found to be hydrolyzed to FMN and AMP by a plasma enzyme, the activity of which in normal adults being estimated as 32.6+9.1 m,uM/ml plasma/min and AMP, thus produced, and/or FAD might compete for the receptor sites on the platelet surface. This could be the mechanism of the inhibitory effect of FAD upon the platelet functions. The intravenous drip infusion of FAD in a dose of 0.2 mg/kg within 30 min was followed by no marked decrease of platelet aggregation, probably because the plasma concentration of FAD and/or AMP did not reach the levels high enough to suppress the platelet functions.-------FAD; AMP; platelet aggregation; platelet ATP release; FAD hydrolyzing enzyme activity of blood plasma; 1-14C-arachidonic acid metabolism Previously flavine adenine dinucleotide (FAD) has been shown to have an inhibitory effect on the platelet aggregation in vitro (Clayton et al. 1963;Kuzuya 1977;Higashi et al. 1978;Shimada et al. 1980). Such an inhibitory effect of FAD on the platelet function could be demonstrated not only in vitro but also in vivo, when a large dose of FAD was administered intravenously (Shimada et al. 1980).The present investigation was conducted to study the effect of FAD upon the adenosine triphosphate (ATP) release of platelets, the mechanism of action of FAD and the in vivo effect of FAD in an ordinary dose upon the platelet functions. MATERIALS AND METHODSBlood was obtained from normal adult volunteers.Methods were similar to those described in the previous paper (Shimada et al. 1980) unless otherwise indicated.The platelet aggregation and ATP release were measured simultaneously by the use of the Lumi-aggregometer (Chrono-Log Corporation, Havortown, Pa). The citrated platelet-rich plasma (PRP) was preincubated with or without FAD of various concentrations for 20 min, and then the aggregating agent, collagen (HormonChemie, Munchen) or arachidonic acid (Silver et al. 1973) was added.For the study of arachidonic acid-induced platelet aggregation, both PRP and washed platelet suspensions in 0.15 M Tris HCl buffer, pH 7.4 (Okuma 1976) were used.

show abstract

Hydrolysis of flavin-adenine dinucleotide by rat liver lysosomes

Cited by 23 publications

References 17 publications

Flavin Adenine Dinucleotide and Flavin Mononucleotide Metabolism in Rat Liver

Flavin Adenine Dinucleotide and Flavin Mononucleotide Metabolism in Rat Liver

A Regulatory Role of NAD Redox Status on Flavin Cofactor Homeostasis inS. cerevisiaeMitochondria

Further studies on the effects of flavine adenine dinucleotide on the platelet functions.

Contact Info

Product

Resources

About