Microbial transformations of N-(4-chlorphenyl)-benzoisothiazolone

Summary The microbial degradation of N-(4-chlorphenyl)-benzoisothiazolone (1) was studied by Streptomyces species in analogy to the metabolism of this drug in animals. As main metabolite 2-thiomethyl-N-(4-chlorphenyl)-benzamide (3) was found. The corresponding sulfoxide (4) and the sulfone (5) were obtained as further transformation products. The unmethylated 2-sulfhydryl compound (2) could be isolated in only small amounts. The degradation pathway follows by these results via a reductive cleavage of the S-N-bond of the isothiazolone ring with subsequent methylation of the sulfhydryl group and further oxidation of the resulting thiomethyl substituent.     

Testosterone metabolism by cytochrome P-450 isozymes RLM3 and RLM5 and by microsomes. Metabolite identification

Testosterone metabolism by cytochrome P-450 isozymes RLM3 and RLM5 in a reconstituted system and by rat liver microsomes was examined. Eleven metabolites were detected. Two of these, found in spots 2 and 4 of a thin layer plate, were only formed by the rat liver microsomes and may represent reductive metabolites of testosterone. A number of monohydroxy metabolites were conclusively identified by gas chromatography-mass spectrometry. These include the 2-, 6 beta-, 7 alpha-, and 16 alpha-hydroxy isomers. Liver microsomes formed the 2 alpha- and 2 beta-epimers in a 1:2 ratio and both co-chromatographed with a third reduced metabolite in thin layer plate spot 4. In contrast with RLM5 about 90% of the 2-hydroxy isomer was the 2 alpha-epimer. RLM3 did not perform the 2-hydroxylation in detectable amounts. The 6 beta-isomer was a major metabolite of RLM3 and microsomes, but a minor product of metabolism by RLM5. In contrast, the 7 alpha-isomer was a minor metabolite of RLM3, was not formed by RLM5, and was a major microsomal metabolite. Hydroxylation at position 16 alpha was a major activity of RLM5 and the heterogeneous microsomal cytochromes, but with RLM3 it was a minor reaction. One new metabolite was found which appeared to be hydroxylated in the D-ring, had a mass spectrum different from both 16 alpha- and 16 beta-hydroxytestosterone, and was tentatively identified as a 15-hydroxy isomer. In agreement with the literature, androstene-3,17-dione was found to be an oxidative metabolite of testosterone by both microsomes and purified cytochrome P-450. It was a major metabolite of RLM5 but was not produced by RLM3. Studies with 18O2 and H218O conclusively show that oxidation of testosterone at C-17 does not involve transient incorporation of an oxygen atom in this position. A mechanism is suggested whereby cytochrome P-450 acts as a peroxidase in the formation of androstenedione.     

Effects of oxygen deficiency on survival and glycogen content of Chironomus anthracinus (Diptera,Chironomidae) under laboratory and field conditions

Growth and glycogen content of Chironomus anthracinus in Lake Esrom, Denmark was examined during summer stratification in 1992 and 1993. Simultaneously, effects of oxygen deficiency on glycogen utilization and survival were experimentally studied. The population consisted of almost fullgrown 4th instar larvae in 1992 and 2nd and 3rd instar larvae in 1993. Growth rate and glycogen content changed as hypolimnetic oxygen deficiency increased. During a 1st phase of stratification dry weight and glycogen content increased (2nd and 3rd instars) or was almost constant (4th instar) but decreased significantly during the following 2nd phase. This change from growth to degrowth and utilization of endogenous glycogen reserves correlated with a change in the thickness of the microxic layer (<0.2 mg O₂ 1^–1) above the sediment surface. The layer increased from 2–3 m in phase 1 to 4–5 m in phase 2, and we suggest that this deteriorated the oxygen conditions and resulted in a change in larval energy metabolism from fully aerobic during the 1st phase to partly anaerobic in the 2nd phase. During the 2nd phase larval metabolism was estimated at less than 20% of normoxic rate. Experimental exposure of the larvae to anoxia indicated highly different survival of young larvae (2nd and 3rd instars) and older larvae (large 4th instars). The morality of young larvae was 50% after three days in anoxia at 10 °C, whereas only 25% of the older larvae had died after 3–4 weeks under similar conditions. Extending the treatment, however, resulted in increased death rate of the 4th instar larvae with only 10% surviving after seven weeks. The anaerobic metabolism of 4th instar larvae as estimated from glycogen degradation at 10 °C was 5% of normoxia in the interval from 0–5 days but 1.5% in the interval from 20–25 days. It is concluded that survival of C. anthracinus in anoxia is very limited, but traces of oxygen in the environment allowing for faint aerobic metabolism prolong the survival time of the larvae from a few days (2nd and 3rd instars) or a few weeks (4th instar) to probably 3–4 months.     

Effect of cyclic AMP-dependent phosphorylation on the reactivity of sulphydryl groups in cardiac sarcoplasmic reticulum.

1. The metabolism of 4-ethynylbiphenyl has been studied in vitro with subcellular fractions of normal and induced rat liver, and rat intestinal microflora (caecal contents). 2. Oxidation was NADPH-dependent, was inhibited by CO and stimulated by pretreatment with phenobarbitone or 3-methylcholanthrene. 3. Oxidation of the ethynyl group occurred in washed microsomal preparations, but not significantly in soluble fractions. Oxidation of the ethynyl group by a microsomal fraction preceded aromatic hydroxylation and no metabolites containing the intact ethynyl group were detected. 4. The major metabolite in liver fractions was biphenyl-4-ylacetic acid. This was the only product produced by a modified Udenfriend system. 5. Metabolism of 4-ethynylbiphenyl by rat caecal contents under anaerobic conditions produced very small amounts of 4-vinylbiphenyl. 6. In a modified Ames test with Salmonella typhimurium TA98, 4-ethynylbiphenyl gave a weak positive result that was doubled after 'activation' with an induced rat S9 fraction.     

In vitro metabolism of aflatoxin B1 by larvae of navel orangeworm, Amyelois transitella (Walker) (Insecta, Lepidoptera, Pyralidae) and codling moth, Cydia pomonella (L.) (Insecta, Lepidoptera, Tortricidae)

Larvae of the navel orangeworm (NOW), Amyelois transitella (Walker), a major pest of almonds and pistachios, and the codling moth (CM), Cydia pomonella (L.), the principal pest of walnuts and pome fruits, are commonly found in tree nut kernels that can be contaminated with aflatoxin, a potent carcinogen. The ability of larvae of these insects to metabolize aflatoxin B1 (AFB1) was examined. A field strain of NOW produced three AFB1 biotransformation products, chiefly aflatoxicol (AFL), and minor amounts of aflatoxin B2a (AFB2a) and aflatoxin M1 (AFM1). With AFL as a substrate, NOW larvae produced AFB1 and aflatoxicol M1 (AFLM1). A lab strain of CM larvae produced no detectable levels of AFB1 biotransformation products in comparison to a field strain which produced trace amounts of only AFL. Neither NOW nor CM produced AFB1-8,9-epoxide (AFBO), the principal carcinogenic metabolite of AFB1. In comparison, metabolism of AFB1 by chicken liver yielded mainly AFL, whereas mouse liver produced mostly AFM1 at a rate eightfold greater than AFL. Mouse liver also produced AFBO. The relatively high production of AFL by NOW compared to CM may reflect an adaptation to detoxify AFB1. NOW larvae frequently inhabit environments highly contaminated with fungi and, hence, aflatoxin. Only low amounts, if any, of this mycotoxin occur in the chief CM hosts, walnuts, and pome fruits. Characterizations of enzymes and co-factors involved in biotransformation of AFB1 are discussed.     

In vitro metabolism of the pro-carcinogen aflatoxin B1 by liver preparations of the calf, nurse shark and clearnose skate

1. Liver postmitochondrial supernatant preparations of calf, clearnose skate, and nurse shark were able to metabolize the fungal toxin aflatoxin B1 to various metabolites. 2. Calf liver produced aflatoxin M1 and Q1 as the major chloroform soluble metabolites, with small amounts of aflatoxicol formed during incubation. 3. Liver preparations of the elasmobranchs, however, produced aflatoxicol as the major chloroform soluble metabolite with no other metabolite being detected. 4. The water soluble metabolite profiles for the three species were also quite different with the tris diol adduct being produced to a much greater extent in calf liver preparations. 5. Aflatoxicol production by the elasmobranch liver homogenates was reversible with the skate reconverting a large amount (30%) of aflatoxicol to AFB1. The nurse shark, however, appeared to convert a portion of aflatoxicol to an unknown metabolite more polar than AFB1. 6. Calf liver DNA bound approximately 3 x more 3H-AFB1 than shark liver DNA.     

Analysis of polyunsaturated fatty acid composition of Strongyloides ratti in relation to development

The effect of linolenic acid (C18:3 omega 3) on the development of Strongyloides ratti first-stage larvae (L1) in culture was studied. The fatty acid composition of S. ratti free-living generations was analyzed by gas chromatography. L1 had abundant linoleic acid (C18:2 omega 6) but its proportion decreased with development. On the contrary, eicosapentaenoic acid (C20:5 omega 3) and C20:4 omega 3 were prominent in the filariform larva (L3). Because C20:5 omega 3 is generally synthesized from C18:3 omega 3 via C20:4 omega 3, the high ratio of C20:5 omega 3/C18:3 omega 3 of L3 in all the free-living generations suggested that polyunsaturated fatty acid metabolism, particularly the omega-3 series, and eicosanoids produced had important roles in the development of S. ratti L1.     

Susceptibility of <Emphasis Type="Italic">Amblyomma variegatum</Emphasis> ticks to acaricides in Ghana

The susceptibility of unfed and fed stages of larvae, nymphs and adult females of Amblyomma variegatum ticks were tested using Shaws filter paper dip method against four acaricides; chlorfenvinphos and dioxathion, chlorfenvinphos, gamma benzene hexachloride and amitraz at four different concentrations including the recommended dose rates. Based on their lethal concentrations (LC₅₀ & LC₉₀) chlorfenvinphos and dioxathion combined and chlorfenvinphos alone placed first and second, respectively, in all stages except at the unfed nymphal stage where gamma benzene hexachloride topped with a LC₅₀ of 0.001629, while chlorfenvinphos and dioxathion combined and chlorfenvinphos alone had LC₅₀ of 0.001794 and 0.002258, respectively. Amitraz appeared to have a quick knock-down effect on larvae and nymphs but at the recommended dose rate, showed no mortality of the ticks at that stage. However, at a concentration of 0.040%, amitraz showed a 100% inhibition of oviposition and hatching of laid eggs. Gamma benzene hexachloride produced only 66% inhibition of oviposition while chlorfenvinphos and dioxathion combined and chlorfenvinphos alone produced 100% inhibition of oviposition at their recommended dose rates. Fed nymphs were more susceptible than the unfed nymphs. Eggs laid by engorged female ovipositing ticks, applied with gamma benzene hexachloride, hatched.This revised version was published online in May 2005 with a corrected cover date.     

Relative tolerance of cirripede larval stages to acute thermal shock: A laboratory study

(1) Meroplankters drawn into once-through cooling circuits of coastal power plants are subjected to transient thermal stress. The effect of such acute thermal shock on the development of barnacle larvae was studied in the laboratory.

(2) The response of the barnacle larvae (naupliar and cyprid stages) to elevated temperature was dependent on exposure time and their stage of development.

(3) Among the stages tested, N-6 larvae showed maximum tolerance. Exposure to 37°C did not affect larval survival, but delayed development of N-2 larva to cypris by one day.

(4) Exposure at 40°C delayed, hastened or did not affect the development time of N-2 and N-4 larvae through cypris, depending on exposure time.

(5) Ten mins exposure at 43°C proved lethal to all larval stages with mortality ranging from 20 to 86%.

(6) Development success of the surviving larvae, measured in terms of cypris yield, showed no significant difference from controls, at temperatures below 40°C.

(7) Settlement activity was significantly affected in only those cyprid larvae which were exposed to 43°C for 10 min.

(8) Results of the present study indicate that thermal stress experienced in the once-through cooling system does not have significant impact on survival and development of the barnacle larvae at temperatures of 37–40°C.     

Hydroxylation of the herbicide isoproturon by fungi isolated from agricultural soil

Several asco-, basidio-, and zygomycetes isolated from an agricultural field were shown to be able to hydroxylate the phenylurea herbicide isoproturon [N-(4-isopropylphenyl)-N',N'-dimethylurea] to N-(4-(2-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea and N-(4-(1-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea. Bacterial metabolism of isoproturon has previously been shown to proceed by an initial demethylation to N-(4-isopropylphenyl)-N'-methylurea. In soils, however, hydroxylated metabolites have also been detected. In this study we identified fungi as organisms that potentially play a major role in the formation of these hydroxylated metabolites in soils treated with isoproturon. Isolates of Mortierella sp. strain Gr4, Phoma cf. eupyrena Gr61, and Alternaria sp. strain Gr174 hydroxylated isoproturon at the first position of the isopropyl side chain, yielding N-(4-(2-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea, while Mucor sp. strain Gr22 hydroxylated the molecule at the second position, yielding N-(4-(1-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea. Hydroxylation was the dominant mode of isoproturon transformation in these fungi, although some cultures also produced traces of the N-demethylated metabolite N-(4-isopropylphenyl)-N'-methylurea. A basidiomycete isolate produced a mixture of the two hydroxylated and N-demethylated metabolites at low concentrations. Clonostachys sp. strain Gr141 and putative Tetracladium sp. strain Gr57 did not hydroxylate isoproturon but N demethylated the compound to a minor extent. Mortierella sp. strain Gr4 also produced N-(4-(2-hydroxy-1-methylethyl)phenyl)-N'-methylurea, which is the product resulting from combined N demethylation and hydroxylation.     

The cardiac effects of amitraz in the guinea-pig in vivo and in vitro

Intravenous amitraz caused significant hypotension and bradycardia in pentobarbitone anaesthetized guinea-pigs. Depression of blood pressure reached a plateau with a dose of 10 mg/kg but heart rate continued to fall in a dose-dependent manner, up to a fall of 90 beats per minute after a total of 160 mg/kg/min. Amitraz was then tested on spontaneously beating guinea-pig isolated atria. The maximum bath concentration approximated a blood concentration produced by 5 mg/kg amitraz in the guinea-pig (2.3 X 10(-4) M). Amitraz did not significantly shift the dose-response curve to isoprenaline or acetylcholine but antagonized histamine rate responses competitively in the presence of propranolol (2 X 10(-6) M). Propranolol unmasked a dose-dependent depressant effect of amitraz on atrial rate, an effect abolished with atropine (1 X 10(-5) M). Amitraz increased atrial force of contraction, an effect which was not seen when propranolol was present in the bath solution. Amitraz also depressed atrial rate directly, but this effect was minor in comparison to bradycardia seen in the guinea-pig. It is likely that the cardiovascular depression seen in the guinea-pig following amitraz i.v. is caused by an alteration in autonomic drive rather than a significant direct cardiac effect.     

Simultaneous determination of amitraz and its metabolite in human serum by monolithic silica spin column extraction and liquid chromatography-mass spectrometry

A simple, rapid, sensitive, and specific liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the quantification of amitraz and its metabolite in human serum. Both the compounds were extracted using monolithic silica spin columns with acetonitrile. The chromatographic separation was performed on a reverse-phase C(18) column with a mobile phase of 10 mM ammonium formate-acetonitrile. The protonated analyte was quantitated in positive ionization by mass spectrometry. The method was validated over the concentration range of 25-1000 ng/ml for amitraz and its metabolite in human serum. For both compounds, the limit of detection was 5 ng/ml. The method was applied to serum samples taken from an attempted suicide patient, and only small volumes of serum were required for the simultaneous determination of these compounds.     

Studies on anabolic steroids--8. GC/MS characterization of unusual seco acidic metabolites of oxymetholone in human urine.

One of the biotransformation routes of oxymetholone (17 beta-hydroxy-2-hydroxymethylene-17 alpha-methyl-5 alpha-androstan-3-one) in man leads to the formation of 17 beta-hydroxy-17 alpha-methyl-5 alpha-androstan-3-one (mestanolone). To demonstrate that this latter steroid may be formed by decarboxylation of an intermediate metabolite of oxymetholone bearing a 2-carboxylic group, we studied the urinary excretion of oxymetholone acidic metabolites. Five new acidic metabolites are reported here for the first time, among which four are unusual seco steroids resulting from the oxidative cleavage of the A-ring. The most abundant compound is 17 beta-hydroxy-17 alpha-methyl-2,3-seco-5 alpha-androstane-2,3-dioic acid 1, the cumulative excretion of which accounted for 1.52% of the dose. Three other seco diacids were produced in smaller amounts, namely 17 beta-hydroxy-17 alpha-methyl-2,3-seco-5 alpha-androstane-2,4- dicarboxylic acid 3, 17 beta-hydroxy-17 alpha-methyl-1,3-seco-5 alpha-androstane-1,3-dioic acid 4 and 17 beta-hydroxy-17 alpha-methyl-2,4-seco-5 alpha-androstane-2,4-dioic acid 5. The fifth acidic metabolite was identified as 3 alpha, 17 beta-dihydroxy-17 alpha-methyl-5 alpha-androstane-2 beta-carboxylic acid 2. The excretion in urine of these acidic metabolites suggests that the 2-hydroxymethylene group in oxymetholone is readily oxidized to yield the corresponding beta-keto acid which can be (1) decarboxylated to form mestanolone; (2) reduced at C-3 to give compound 2; and (3) further oxidized to afford the unexpected seco diacids 1, 3, 4 and 5. The identity of compounds 1 and 2 was ascertained by GC/MS and 1H and 13C-NMR analysis of reference compounds. The other metabolites were characterized by GC/MS analysis.     

Manifestations of resistance to Haemonchus contortus in sheep: Worm populations and abomasal changes in sheep superinfected with 1,000,000 larvae of H. contortus

The super-infecting dose produced a marked rise in gastric pH in all sheep from the 3rd day after administration of larvae. Expulsion of the existing population of adult worms may have begun on the 4th day but was still only completed in 3/6 sheep on the 5th day. The larvae caused extensive damage in the individual glands which they parasitised. Very few of the 10⁶ larvae survived for 27 days and only in 1/8 sheep had they developed beyond early 4th stage at 27 days. Extensive histological changes were seen in the fundic mucosa beginning as early as 2 days after the superinfection. While the pH change preceded expulsion of the adults and was consistent in its timing, the timing of the expulsion was irregular. This throws doubt on the hypothesis that the change in physico-chemical conditions produced by the superinfecting larvae is the only cause of the expulsion of the adult worms.     

N-15 in vivo NMR spectroscopic investigation of nitrogen deprived cell suspensions of the green alga Chlorella fusca

The possibility to apply N-15 in vivo NMR spectroscopy to study algal N-metabolism has been investigated. N-15 labelled cells of the green alga Chlorella fusca, subjected to nitrogen starvation and N-14 labelled cells supplied with K¹⁵NO₃ after prolonged nitrogen starvation were monitored by N-15 in vivo NMR spectroscopy at different times after the change in their nitrogen supply. During 20–40 min, necessary for the acquisition of 1 spectrum, the cells were under dark anaerobic conditions, but the relative amounts of the metabolites detected did not change. Signals from 2 acid amides, from the side chain nitrogens of arginine and lysine, from prolin as well as 4 signals from α amino groups of amino acids were detected. Besides two signals not yet reported in the literature were found. They may be due to amino compounds, but not to amino acids. The amount of free amino acids in the cells increases not only upon resupply of nitrogen starved cells with nitrate but also during the first hours after nitrate depletion. The spectra obtained from N-15 labelled autospores show that N-15 in vivo NMR spectroscopy can be applied to the investigation of N metabolism of the cells.     

In vitro metabolism of the cardiotonic steroids gomphogenin and calactin

The metabolism of gomphogenin and calactin was studied in vitro using respectively microsomes and the S9 fraction of homogenates from rat liver. These two substrates were previously shown to be in vitro and in vivo metabolites of gomphoside, a cardiotonic steroid belonging to a class of 5 alpha-cardenolide glycosides with doubly-linked hexosulose sugars. Structures of new metabolites were elucidated using 400 MHz 1H-NMR and chemical ionization mass spectrometry, while known compounds were identified by direct comparison. The major metabolite isolated from gomphogenin (2 alpha-hydroxyuzarigenin) metabolism was the oxidation product 2-oxo-uzarigenin which was further oxidized metabolically to 4 alpha-hydroxy-2-oxo-uzarigenin. Other metabolites were 2 alpha-hydroxyuzarigenone and its reduction product 3-epigomphogenin. Calactin was oxidized in vitro to 10-carboxyl-19-norgomphoside, the predominant metabolite, and underwent cleavage of the doubly-linked sugar to yield calotropagenin.     

Importance of multiple hydroxylated metabolites in hexamethylphosphoramide (HMPA)-mediated mutagenesis in Drosophila melanogaster

The mutagenic profiles in Drosophila and the influence of inhibition of metabolism on genotoxic activity were determined for hexamethylphosphoric triamide (HMPA), some synthetically prepared presumed metabolites and ethylated analogs. Demethylated HMPA metabolites are considerably less mutagenic than HMPA, dependent on the degree of demethylation. The mutagenicity of the presumptive primary metabolite, hydroxymethyl pentamethylphosphoramide (HM-Me5-PA), is comparable to HMPA and can be decreased considerably by inhibition of the metabolism by 1-phenylimidazole or iproniazid. This suggests that further oxidative metabolism is required for mutagenic activity. The mutagenicity of the doubly hydroxylated HMPA metabolite, N,N'-bis(hydroxymethyl)-tetramethylphosphoramide (N,N'-(HM)2-Me4-PA) can also be decreased by inhibition of metabolism, whereas the 3-fold hydroxylated N,N',-N"-(HM)3-Me3-PA is not affected by pretreatment with enzyme inhibitors, indicating that no further oxidative metabolism is required for its activation. A second hydroxylation on 1 dimethylamino group, forming N,N-(HM)2-Me4-PA, results in a drastic loss of mutagenic activity. Further oxidation of HM-Me5-PA to formyl pentamethylphosphoramide (formyl-Me5-PA) also leads to a strong reduction of the genotoxic activity. The rearrangement product of N-oxidation, N-[bis(dimethylamino)phosphinyl)-oxy)dimethylamine (HMPOA) is not mutagenic in Drosophila. The very low mutagenicity of hexaethylphosphoramide (Et6-PA) allowed us to study the mutagenicity of some ethyl-hydroxymethyl hybrid compounds. For the ethylated phosphoramides also the presence of only 1 hydroxymethyl group is insufficient for mutagenic activity, whereas the introduction of 2 or 3 hydroxymethyl groups resulted in considerable genotoxicity in the sex-linked recessive lethal (SLRL) test as well as in the ring-X loss test. It is concluded that the bioactivation of HMPA in Drosophila proceeds via multiple metabolic hydroxylations to form multifunctional, cross-linking agents. The presence of an oxygen atom on the phosphorus appears to be a prerequisite for the genotoxic activity of HMPA as hexamethylphosphorus triamide (HMPT), a derivative lacking this oxygen, is only weakly mutagenic in Drosophila. The results presented in this paper do not support the theory that formaldehyde is the active principle of activated HMPA.     

Properties of heparan sulphate and chondroitin sulphate from young and old human aortae

1. gamma-Hexachlorocyclohexane, gamma-pentachlorocyclohexene and delta-pentachlorocyclohexene were converted by houseflies and grass grubs into metabolites that had chromatographic properties identical with those of S-2,4-dichlorophenylglutathione. 2. The metabolism of gamma-hexachlorocyclohexane and the pentachlorocyclohexene isomers was negligible in newly emerged blowflies, but increased over the next 10 days. 3. The metabolism of both gamma-hexachlorocyclohexane and the pentachlorocyclohexene isomers was inhibited by simultaneous dosage with tetrabromophenolphthalein ethyl ester or Bromophenol Blue in both grass grubs and flies, but only the metabolism of pentachlorocyclohexenes in blowflies was stopped by simultaneous dosage with bis-(N-dimethylaminophenyl)methane. NN-Di-n-butyl-p-chlorobenzenesulphonamide had no effect on the metabolism of pentachlorocyclohexenes by blowflies. 4. The use of these inhibitors and colorimetric assays leads to the conclusion that a pentachlorocyclohexene is not a major intermediary metabolite of gamma-hexachlorocyclohexane in these insects.     

Angiopoietin-like Protein 3 Inhibits Lipoprotein Lipase Activity through Enhancing Its Cleavage by Proprotein Convertases

Lipoprotein lipase (LPL)-mediated lipolysis of triglycerides is the first and rate-limiting step in chylomicron/very low density lipoprotein clearance at the luminal surface of the capillaries. Angiopoietin-like protein 3 (ANGPTL3) is shown to inhibit LPL activity and plays important roles in modulating lipoprotein metabolism in vivo. However, the mechanism by which it inhibits LPL activity remains poorly understood. Using cell-based analysis of the interaction between ANGPTL3, furin, proprotein convertase subtilisin/kexin type 5 (PCSK5), paired amino acid converting enzyme-4 (PACE4), and LPL, we demonstrated that the cleavage of LPL by proprotein convertases is an inactivation process, similar to that seen for endothelial lipase cleavage. At physiological concentrations and in the presence of cells, ANGPTL3 is a potent inhibitor of LPL. This action is due to the fact that ANGPTL3 can enhance LPL cleavage by endogenous furin and PACE4 but not by PCSK5. This effect is specific to LPL but not endothelial lipase. Both N- and C-terminal domains of LPL are required for ANGPTL3-enhanced cleavage, and the N-terminal domain of ANGPTL3 is sufficient to exert its effect on LPL cleavage. Moreover, ANGPTL3 enhances LPL cleavage in the presence of either heparan sulfate proteoglycans or glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1). By enhancing LPL cleavage, ANGPTL3 dissociates LPL from the cell surface, inhibiting both the catalytic and noncatalytic functions of LPL. Taken together, our data provide a molecular connection between ANGPTL3, LPL, and proprotein convertases, which may represent a rapid signal communication among different metabolically active tissues to maintain energy homeostasis. These novel findings provide a new paradigm of specific protease-substrate interaction and further improve our knowledge of LPL biology.