The Experts below are selected from a list of 6591 Experts worldwide ranked by ideXlab platform

Radhika Venkataraman - One of the best experts on this subject based on the ideXlab platform.

William C Groutas - One of the best experts on this subject based on the ideXlab platform.

Francis Gauthier - One of the best experts on this subject based on the ideXlab platform.

  • neutrophil elastase proteinase 3 and Cathepsin G as therapeutic tarGets in human diseases
    Pharmacological Reviews, 2010
    Co-Authors: Brice Korkmaz, Dieter E Jenne, Marshall S. Horwitz, Francis Gauthier
    Abstract:

    Polymorphonuclear neutrophils are the first cells recruited to inflammatory sites and form the earliest line of defense aGainst invadinG microorGanisms. Neutrophil elastase, proteinase 3, and Cathepsin G are three hematopoietic serine proteases stored in larGe quantities in neutrophil cytoplasmic azurophilic Granules. They act in combination with reactive oxyGen species to help deGrade enGulfed microorGanisms inside phaGolysosomes. These proteases are also externalized in an active form durinG neutrophil activation at inflammatory sites, thus contributinG to the reGulation of inflammatory and immune responses. As multifunctional proteases, they also play a reGulatory role in noninfectious inflammatory diseases. Mutations in the ELA2/ELANE Gene, encodinG neutrophil elastase, are the cause of human conGenital neutropenia. Neutrophil membrane-bound proteinase 3 serves as an autoantiGen in WeGener Granulomatosis, a systemic autoimmune vasculitis. All three proteases are affected by mutations of the Gene (CTSC) encodinG dipeptidyl peptidase I, a protease required for activation of their proform before storaGe in cytoplasmic Granules. Mutations of CTSC cause Papillon-Lefevre syndrome. Because of their roles in host defense and disease, elastase, proteinase 3, and Cathepsin G are of interest as potential therapeutic tarGets. In this review, we describe the physicochemical functions of these proteases, toward a Goal of better delineatinG their role in human diseases and identifyinG new therapeutic strateGies based on the modulation of their bioavailability and activity. We also describe how nonhuman primate experimental models could assist with testinG the efficacy of proposed therapeutic strateGies.

  • measurinG elastase proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates
    Nature Protocols, 2008
    Co-Authors: Brice Korkmaz, Marielise Jourdan, Sylvie Attucci, Maria A Juliano, Timofey Kalupov, Luiz Juliano, Francis Gauthier
    Abstract:

    MeasurinG elastase, proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates

  • MeasurinG elastase, proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates
    Nature Protocols, 2008
    Co-Authors: Brice Korkmaz, Marielise Jourdan, Sylvie Attucci, Maria A Juliano, Timofey Kalupov, Luiz Juliano, Francis Gauthier
    Abstract:

    The neutrophil serine proteases (NSPs) elastase, proteinase 3 and Cathepsin G are multifunctional proteases involved in pathoGen destruction and the modulation of inflammatory processes. A fraction of secreted NSPs remains bound to the external plasma membrane, where they remain enzymatically active. This protocol describes the spectrofluorometric measurement of NSP activities on neutrophil surfaces usinG hiGhly sensitive Abz-peptidyl-EDDnp fluorescence resonance enerGy transfer (FRET) substrates that fully discriminate between the three human NSPs. We describe FRET substrate synthesis, neutrophil purification and handlinG, and kinetic experiments on quiescent and activated cells. These are used to measure subnanomolar concentrations of membrane-bound or free NSPs in low-bindinG microplates and to quantify the activities of individual proteases in bioloGical fluids like expectorations and bronchoalveolar lavaGes. The whole procedure, includinG neutrophil purification and kinetic measurements, can be done in 4–5 h and should not be lonGer because of the lifetime of neutrophils. UsinG this protocol will help identify the contributions of individual NSPs to the development of inflammatory diseases and may reveal these proteases to be tarGets for therapeutic inhibitors.

  • neutrophil elastase proteinase 3 and Cathepsin G physicochemical properties activity and physiopatholoGical functions
    Biochimie, 2008
    Co-Authors: Brice Korkmaz, Thierry Moreau, Francis Gauthier
    Abstract:

    Polymorphonuclear neutrophils form a primary line of defense aGainst bacterial infections usinG complementary oxidative and non-oxidative pathways to destroy phaGocytized pathoGens. The three serine proteases elastase, proteinase 3 and Cathepsin G, are major components of the neutrophil primary Granules that participate in the non-oxidative pathway of intracellular pathoGen destruction. Neutrophil activation and deGranulation results in the release of these proteases into the extracellular medium as proteolytically active enzymes, part of them remaininG exposed at the cell surface. Extracellular neutrophil serine proteases also help kill bacteria and are involved in the deGradation of extracellular matrix components durinG acute and chronic inflammation. But they are also important as specific reGulators of the immune response, controllinG cellular siGnalinG throuGh the processinG of chemokines, modulatinG the cytokine network, and activatinG specific cell surface receptors. Neutrophil serine proteases are also involved in the pathoGenicity of a variety of human diseases. This review focuses on the structural and functional properties of these proteases that may explain their specific bioloGical roles, and facilitate their use as molecular tarGets for new therapeutic strateGies.

Brice Korkmaz - One of the best experts on this subject based on the ideXlab platform.

  • neutrophil elastase proteinase 3 and Cathepsin G as therapeutic tarGets in human diseases
    Pharmacological Reviews, 2010
    Co-Authors: Brice Korkmaz, Dieter E Jenne, Marshall S. Horwitz, Francis Gauthier
    Abstract:

    Polymorphonuclear neutrophils are the first cells recruited to inflammatory sites and form the earliest line of defense aGainst invadinG microorGanisms. Neutrophil elastase, proteinase 3, and Cathepsin G are three hematopoietic serine proteases stored in larGe quantities in neutrophil cytoplasmic azurophilic Granules. They act in combination with reactive oxyGen species to help deGrade enGulfed microorGanisms inside phaGolysosomes. These proteases are also externalized in an active form durinG neutrophil activation at inflammatory sites, thus contributinG to the reGulation of inflammatory and immune responses. As multifunctional proteases, they also play a reGulatory role in noninfectious inflammatory diseases. Mutations in the ELA2/ELANE Gene, encodinG neutrophil elastase, are the cause of human conGenital neutropenia. Neutrophil membrane-bound proteinase 3 serves as an autoantiGen in WeGener Granulomatosis, a systemic autoimmune vasculitis. All three proteases are affected by mutations of the Gene (CTSC) encodinG dipeptidyl peptidase I, a protease required for activation of their proform before storaGe in cytoplasmic Granules. Mutations of CTSC cause Papillon-Lefevre syndrome. Because of their roles in host defense and disease, elastase, proteinase 3, and Cathepsin G are of interest as potential therapeutic tarGets. In this review, we describe the physicochemical functions of these proteases, toward a Goal of better delineatinG their role in human diseases and identifyinG new therapeutic strateGies based on the modulation of their bioavailability and activity. We also describe how nonhuman primate experimental models could assist with testinG the efficacy of proposed therapeutic strateGies.

  • measurinG elastase proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates
    Nature Protocols, 2008
    Co-Authors: Brice Korkmaz, Marielise Jourdan, Sylvie Attucci, Maria A Juliano, Timofey Kalupov, Luiz Juliano, Francis Gauthier
    Abstract:

    MeasurinG elastase, proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates

  • MeasurinG elastase, proteinase 3 and Cathepsin G activities at the surface of human neutrophils with fluorescence resonance enerGy transfer substrates
    Nature Protocols, 2008
    Co-Authors: Brice Korkmaz, Marielise Jourdan, Sylvie Attucci, Maria A Juliano, Timofey Kalupov, Luiz Juliano, Francis Gauthier
    Abstract:

    The neutrophil serine proteases (NSPs) elastase, proteinase 3 and Cathepsin G are multifunctional proteases involved in pathoGen destruction and the modulation of inflammatory processes. A fraction of secreted NSPs remains bound to the external plasma membrane, where they remain enzymatically active. This protocol describes the spectrofluorometric measurement of NSP activities on neutrophil surfaces usinG hiGhly sensitive Abz-peptidyl-EDDnp fluorescence resonance enerGy transfer (FRET) substrates that fully discriminate between the three human NSPs. We describe FRET substrate synthesis, neutrophil purification and handlinG, and kinetic experiments on quiescent and activated cells. These are used to measure subnanomolar concentrations of membrane-bound or free NSPs in low-bindinG microplates and to quantify the activities of individual proteases in bioloGical fluids like expectorations and bronchoalveolar lavaGes. The whole procedure, includinG neutrophil purification and kinetic measurements, can be done in 4–5 h and should not be lonGer because of the lifetime of neutrophils. UsinG this protocol will help identify the contributions of individual NSPs to the development of inflammatory diseases and may reveal these proteases to be tarGets for therapeutic inhibitors.

  • neutrophil elastase proteinase 3 and Cathepsin G physicochemical properties activity and physiopatholoGical functions
    Biochimie, 2008
    Co-Authors: Brice Korkmaz, Thierry Moreau, Francis Gauthier
    Abstract:

    Polymorphonuclear neutrophils form a primary line of defense aGainst bacterial infections usinG complementary oxidative and non-oxidative pathways to destroy phaGocytized pathoGens. The three serine proteases elastase, proteinase 3 and Cathepsin G, are major components of the neutrophil primary Granules that participate in the non-oxidative pathway of intracellular pathoGen destruction. Neutrophil activation and deGranulation results in the release of these proteases into the extracellular medium as proteolytically active enzymes, part of them remaininG exposed at the cell surface. Extracellular neutrophil serine proteases also help kill bacteria and are involved in the deGradation of extracellular matrix components durinG acute and chronic inflammation. But they are also important as specific reGulators of the immune response, controllinG cellular siGnalinG throuGh the processinG of chemokines, modulatinG the cytokine network, and activatinG specific cell surface receptors. Neutrophil serine proteases are also involved in the pathoGenicity of a variety of human diseases. This review focuses on the structural and functional properties of these proteases that may explain their specific bioloGical roles, and facilitate their use as molecular tarGets for new therapeutic strateGies.

Krzysztf Rolka - One of the best experts on this subject based on the ideXlab platform.

  • selection of peptomeric inhibitors of bovine α chymotrypsin and Cathepsin G based on trypsin inhibitor sfti 1 usinG a combinatorial chemistry approach
    Molecular Diversity, 2010
    Co-Authors: Anna łegowska, Adam Lesner, Magdalena Wysocka, Dawid Debowski, Krzysztf Rolka
    Abstract:

    A peptomeric library consistinG of 360 monocyclic analoGues of trypsin inhibitor SFTI-1 isolated from sunflower seeds was desiGned and synthesized by a solid-phase approach in order to select chymotrypsin and Cathepsin G inhibitors. All peptomers contained a proteinoGenic-Phe-mimickinG N-benzylGlycine (Nphe) at positions 5 and 12. Into the synthesized library, different peptoid monomers were introduced in the 7–10 seGment. Deconvolution of the library aGainst both proteinases throuGh an iterative method in solution revealed that the stronGest chymotrypsin inhibitory activity was displayed by two analoGues, [Nphe5,12]SFTI-1 (1) and [Nphe5,12, Naem8]SFTI-1 (2), where Naem stands for N-(2-morpholinoethyl)Glycine. After deconvolution aGainst a Cathepsin G analoGue, [Nphe5,12, Npip8,9, Nnle10] SFTI-1 (3) (Npip = N-(3,4-methylenedioxybenzyl)Glycine) appeared to be the most potent inhibitor with a hiGh serum stability. It is worth notinG that the analoGues obtained by a combinatorial approach display hiGh specificity towards one of the experimental enzymes. Another interestinG feature is the lack of Pro8 in analoGues 2 and 3, the amino acid residue absolutely conserved in the family of Bownan–Birk inhibitors.

  • introduction of non natural amino acid residues into the substrate specific p1 position of trypsin inhibitor sfti 1 yields potent chymotrypsin and Cathepsin G inhibitors
    Bioorganic & Medicinal Chemistry, 2009
    Co-Authors: Anna łegowska, Adam Lesner, Magdalena Wysocka, Dawid Debowski, Krzysztf Rolka
    Abstract:

    Abstract A series of trypsin inhibitor SFTI-1compounds modified in substrate-specific P1 position was synthesized by the solid-phase method. Lys5 present in the wild inhibitor was replaced by Phe derivatives substituted in para position of the phenyl rinG, l -pyridylalanine and N-4-nitrobenzylGycine. Their inhibitory activities with bovine α-chymotrypsin and Cathepsin G were estimated by determination of association equilibrium constants (Ka). All analoGues inhibited bovine α-chymotrypsin. The hiGhest inihbitory activity displayed peptides with the fluorine, nitro and methyl substituents. They were 13–15-fold more active than [Phe5]SFTI-1 used as a reference. They are the most potent chymotrypsin inhibitors of this size. Substitution of Lys5 by Phe did not chanGe the Cathepsin G inhibitory activity. Introduction of Phe(p-F), Phe(p-NH2) and Phe(p-CH3) in this position retained the affinity towards this proteinase, whereas Phe(p-Guanidine) Gave an inhibitor more than twice as active, which appeared to be stable in human serum. On the other hand, a peptomeric analoGue with N-4-nitrobenzylGlycine failed to inhibit Cathepsin G. Despite the fact the introduced amino acids were non-coded, the peptide bonds formed by them were hydrolyzed by chymotrypsin. We postulate that additional interaction of para-substitutents with the enzyme are responsible for the enhanced inhibitory activity of the analoGues.

  • New chromoGenic substrates of human neutrophil Cathepsin G containinG non-natural aromatic amino acid residues in position P_1 selected by combinatorial chemistry methods
    Molecular Diversity, 2007
    Co-Authors: Magdalena Wysocka, Elżbieta Bulak, Hanna Miecznikowska, Anna Jaśkiewicz, Adam Lesner, Anna Łęgowska, Krzysztf Rolka
    Abstract:

    Specificity of human Cathepsin G was explored usinG combinatorial chemistry methods. Deconvolution of a tetrapeptide library, where 5-amino-2-nitrobenzoic acid served as a chromophore attached at the C-terminus, yielded the active sequence Phe-Val-Thr-Tyr-Anb^5,2-NH_2. This sequence was used for a second-Generation library with the General formula Ac-Phe-Val-Thr-X-Anb^5,2-NH_2, where position X was replaced with several amino acids: l -pyridyl- alanine (Pal), 4-nitro- l -phenylalanine (Nif), 4-amino- l - phenylalanine (Amf), 4-carboxy- l -phenylalanine (Cbf), 4-Guanidine- l -phenylalanine (Gnf), 4-methyloxycarbonyl- l -phenylalanine (Mcf), 4-cyano- l -phenylalanine (Cyf), Phe, Tyr, ArG and Lys. Specificity liGand parameters, k _cat and K _M, with human Cathepsin G were determined for all chromoGenic substrates synthesized. The hiGhest value of the specificity constant ( k _cat/ K _M) was obtained for a substrate with the Gnf residue in position P_1. This peptide was 10 times more active than the second most active substrate which contained the Amf residue. The followinG order of potency was established: Gnf >  > Amf > Tyr = Phe > ArG= Lys > Cyf. Substrate specificity for Cathepsin G is Greatly enhanced when an aromatic side chain and a stronG positive charGe are incorporated in residue P_1.