Inhibition by aminoacyl‐chloromethane protease inhibitors of superoxide anion production by phorbol‐ester‐stimulated human neutrophils

Conseiller, Emmdniiei C.; Schott, D.; Lederer, Florence

doi:10.1111/j.1432-1033.1990.tb19344.x

Cited by 6 publications

(19 citation statements)

References 61 publications

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“…[3HITosPheCH,Cl was obtained as described in [49]. The specific activity of the radiolabel was 3 Ci/mmol.…”

Section: Treatment Of Neutrophils With [3hltosphechc1 and Analysis Omentioning

confidence: 99%

“…In the cell-free assay, the superoxide-producing activity of membranes and cytosol prepared from inhibited cells is unimpaired Conseiller et al [49] found that a homologous combination of cytosol and membranes from cells treated with TosPheCH,Cl (but not PMA) showed a decreased superoxide production in the cell-free activation system in the presence of SDS. Heterologous recombinations (i.e.…”

Section: Particulate Nadph Oxidase Prepared From Tosphechci-treated mentioning

confidence: 99%

“…Indeed, Conseiller et al [49] separated membranes and cytosol by a 100000 X g ultracentrifugation of a post-nuclear supernatant and used approximately equivalent amounts of proteins from the two fractions, whereas we introduced a number of improvements suggested by the literature (removal of granules by a 10000 X g centrifugation [52], use of a higher ratio of cytosolic/membrane proteins [53,56,571, adjustment of the arachidonate concentration [53] and addition of GTP[S] [25]). Thus the specific activities obtained in our studies were approximately tenfold higher than those of the system used by Conseiller et al [49] when values were calculated on a cell-equivalent basis, and from 20-fold to more than 100-fold for valuedmg membrane protein. Our specific activities (Fig.…”

Section: Particulate Nadph Oxidase Prepared From Tosphechci-treated mentioning

confidence: 99%

“…More recently, carbobenzoxy-leucyl-tyrosyl chloromethane (zLeuTyrCH,CI) was found to inhibit the respiratory burst elicited by Met-Leu-Phe, but not that triggered by PMA [42,431. In contrast, variable results were obtained with phenylmethanesulfonyl fluoride (PheMeS0,F) and diisopropylfluorophosphate, from no effect to potent inhibition, depending on the reagent concentration and the cell stimulus [29,30,33,36,[39][40][41][44][45][46][47][48][49]. Many of these studies concluded that a protease was involved in NADPH oxidase activation.…”

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confidence: 99%

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Aminoacyl chloromethanes as tools to study the requirements of NADPH oxidase activation in human neutrophils

Chollet-Przednowed

Lederer

1993

European Journal of Biochemistry

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Previous studies from this laboratory described the kinetic characteristics of the inhibition by tosylphenylalanine chloromethane (TosPheCH,Cl) on superoxide anion production by human neutrophils (PMN) stimulated with a phorbol ester (PMA). In this study we present further evidence concerning the potential role of the chloromethane target in the normal cellular activation of NADPH oxidase. When PMN are treated with TosPheCH,Cl and subsequently PMA, or with the two reagents in the reverse order, the inhibition of superoxide production by the intact cells is still present in a particulate NADPH oxidase fraction prepared from these cells. Nevertheless, when cells incubated only with the chloromethane and not with PMA are disrupted, both their cytosolic and membrane fractions are fully competent in the cell-free activation assay. Thus, the chloromethane target has a role in NADPH oxidase activation exclusively at the cellular level. This observation constitutes additional evidence in favour of the idea that activation in the cell-free system reflects only partially the events which occur in the cells.When cells are activated with PMA, their cytosol displays a loss of activating capacity in the cell-free activation assay in the presence of arachidonate, as was shown before with SDS as activatorBiol. Chem. 265,924-9301. This phenomenon was shown to arise most probably from the translocation of cytosolic factors to the membrane, resulting in a depleted cytosol. When superoxide production was inhibited by cell treatment with TosPheCH,Cl, either before or after activation with PMA, the cytosol from inhibited cells showed a recovery of activation capacity in the cell-free system. This effect probably results from TosPheCH,Cl inhibiting the translocation of the cytosolic factors when added before PMA. This results in an insufficient activation at the membrane level, which was previously considered as an inhibition. The effect of TosPheCH,Cl, when added after PMA, can best be explained again as an inhibition of translocation in the frame of the continuous replenishment In this study, we have also found that [3H]TosPheCH,C1-labeled cells show a prominent radiolabeled band of M, 15000 after SDSPAGE analysis of the cytosolic proteins from inhibited cells; this species has the same migration as the labeled membrane protein detected previously, but appears much more abundant on a proteidcell basis. The potential role of the TosPheCH,Cl protein target is discussed.Polymorphonuclear leukocytes (PMN) constitute the first line of defense against infection in higher organisms. Contact between bacteria o r other pathogens and PMN induces phagocytosis and destruction of the invader cells in the phagolysosomes. An important element of this defense mechanism is constituted by the toxic oxygen derivatives generated as the result of superoxide anion production by NADPH oxidase, an enzyme system which is activated upon cell stimulation [l-31.The best identified redox component of NADPH oxidase, a plasma-membrane bound complex, is the hetero...

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“…[3HITosPheCH,Cl was obtained as described in [49]. The specific activity of the radiolabel was 3 Ci/mmol.…”

Section: Treatment Of Neutrophils With [3hltosphechc1 and Analysis Omentioning

confidence: 99%

Section: Particulate Nadph Oxidase Prepared From Tosphechci-treated mentioning

confidence: 99%

Section: Particulate Nadph Oxidase Prepared From Tosphechci-treated mentioning

confidence: 99%

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confidence: 99%

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Aminoacyl chloromethanes as tools to study the requirements of NADPH oxidase activation in human neutrophils

Chollet-Przednowed

Lederer

1993

European Journal of Biochemistry

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“…The potent inhibitory effect of tosylphenylalanine chloromethane and tosylphenyllysine chloromethane on the respiratory burst led to the idea that a chymotryptic-like protease might play a role in oxidase activation (Conseiller and Lederer, 1989). It turned out that the molecular target of the chloromethane derivative is a 15-kDa hydrophobic protein residing in the plasma membrane of neutrophils, and that the chemical modification of this membrane-bound protein is probably responsible for inactivation of the oxidase (Conseiller et al, 1990).…”

Section: Role Of Activated Protein Kinase C In the Respiratory Burstmentioning

confidence: 99%

The superoxide‐generating oxidase of phagocytic cells

Morel¹,

Doussière²,

Vignais³

1991

European Journal of Biochemistry

537

260

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Professional phagocytes (neutrophils, eosinophils, monocytes and macrophages) possess an enzymatic complex, the NADPH oxidase, which is able to catalyze the one-electron reduction of molecular oxygen to superoxide, 0;. The NADPH oxidase is dormant in non-activated phagocytes. It is suddenly activated upon exposure of phagocytes to the appropriate stimuli and thereby contributes to the microbicidal activity of these cells. Oxidase activation in phagocytes involves the assembly, in the plasma membrane, of membrane-bound and cytosolic components of the oxidase complex, which were disassembled in the resting state. One of the membrane-bound components in resting phagocytes has been identified as a low-potential b-type cytochrome, a heterodimer composed of two subunits of 22-kDa and 91-kDa. The link between NADPH and cytochrome b is probably a flavoprotein whose subcellular localization in resting phagocytes remains to be determined. Genetic defects in the cytochrome b subunits and in the cytosolic factors have been shown to be the molecular basis of chronic granulomatous disease, a group of inherited disorders in the host defense, characterized by severe, recurrent bacterial and fungal infections in which phagocytic cells fail to generate 0 , upon stimulation. The present review is focused on recent data concerning the signaling pathway which leads to oxidase activation, including specific receptors, the production of second messengers, the organization of the oxidase complex and the molecular defects responsible for granulomatous disease.

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The effect of serine and thiol protease inhibitors on the chemiluminescence of human neutrophils in investigations In Vitro

Kantorski¹,

Tchórzewski²

1992

J. Biolumin. Chemilumin.

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We have studied an indirect role of serine and thiol proteases in the activation of human neutrophils in vitro. Stimulation was evaluated using a chemiluminescence (CL) generation system. Receptor-dependent and receptor-independent stimuli were studied, e.g. opsonized zymosan, formyl-methionyl-leucyl-phenylalanine, platelet activating factor, phorbol myristate acetate, and calcium ionophore A23187. The serine protease inhibitors TPCK and TLCK, and thiol protease inhibitor PHMB, diminished the CL with different potencies and in a dose-dependent manner after treatment of cells with the various stimuli. Non-specific serine protease inhibitor, PMSF, and trypsin substrate TAME, showed a low inhibitory potency with respect to CL generation. Synthetic substrates for chymotrypsin (BTEE, ATEE) significantly inhibited CL with the various stimuli used with some differences in susceptibility to their inhibition. Specific chymotrypsin inhibitors diminished both the resting and activator-induced CL. We suggest that cell-bound chymotrypsin-like protease(s) is involved in the activation of signal transduction in human neutrophils after both receptor-dependent and receptor-independent stimulation.

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Inhibition by aminoacyl‐chloromethane protease inhibitors of superoxide anion production by phorbol‐ester‐stimulated human neutrophils

Cited by 6 publications

References 61 publications

Aminoacyl chloromethanes as tools to study the requirements of NADPH oxidase activation in human neutrophils

Aminoacyl chloromethanes as tools to study the requirements of NADPH oxidase activation in human neutrophils

The superoxide‐generating oxidase of phagocytic cells

The effect of serine and thiol protease inhibitors on the chemiluminescence of human neutrophils in investigations In Vitro

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