Mechanistic Studies on the Inactivation of Papain by Epoxysuccinyl Inhibitors

Meara, Joseph P.; Rich, Daniel H.

doi:10.1021/jm950445b

Cited by 75 publications

(101 citation statements)

References 62 publications

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“…Tissues were lysed in a solution containing 50 mM sodium acetate (pH 5.5), 1 mM EDTA, 0.1% CHAPS {3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate}, 0.5% NP-40, 0.1 mg of Pefabloc/ml, and 5 g of pepstatin A/ml (RIPA buffer) for 1 h at 4°C. Equal amounts of lysate from each tissue were labeled with 125 I-JPM565 (20,28) for 1 h at 37°C and then diluted in SDS sample buffer and boiled for 10 min. Samples were fractionated by SDS-PAGE (12.5% gel), dried, and visualized by autoradiography.…”

Section: Methodsmentioning

confidence: 99%

Murine Cathepsin F Deficiency Causes Neuronal Lipofuscinosis and Late-Onset Neurological Disease

Tang

Lee

Galvez

et al. 2006

Molecular and Cellular Biology

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Cathepsin F (cat F) is a widely expressed lysosomal cysteine protease whose in vivo role is unknown. To address this issue, mice deficient in cat F were generated via homologous recombination. Although cat F−/− mice appeared healthy and reproduced normally, they developed progressive hind leg weakness and decline in motor coordination at 12 to 16 months of age, followed by significant weight loss and death within 6 months. cat F was found to be expressed throughout the central nervous system (CNS). cat F−/− neurons accumulated eosinophilic granules that had features typical of lysosomal lipofuscin by electron microscopy. Large amounts of autofluorescent lipofuscin, characteristic of the neurodegenerative disease neuronal ceroid lipofuscinosis (NCL), accumulated throughout the CNS but not in visceral organs, beginning as early as 6 weeks of age. Pronounced gliosis, an indicator of neuronal stress and neurodegeneration, was also apparent in older cat F−/− mice. cat F is the only cysteine cathepsin whose inactivation alone causes a lysosomal storage defect and progressive neurological features in mice. The late onset suggests that this gene may be a candidate for adult-onset NCL.

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Section: Methodsmentioning

confidence: 99%

Murine Cathepsin F Deficiency Causes Neuronal Lipofuscinosis and Late-Onset Neurological Disease

Tang

Lee

Galvez

et al. 2006

Molecular and Cellular Biology

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“…T h e f l u o r e s c e n t S -n i t r o s o p r o b e ( S -n i t r o s o -5 -dimethylaminonaphthalene-1-sulfonyl) was used to demonstrate that the NO donor was bound to the protein as the Cys-S-R complex 91 . Papain as a cysteine protease shares many features of the important mammalian cysteine proteases cathepsin B, H, L and S and the calpains 92 . It should be noted that cysteine proteases are involved in cellular apoptosis, and importantly several members of this group (caspases 1,2,3,4,6,7 and 8) have been shown to reversibly inactivated by NO 93 .…”

Section: Protein Cysteine S-nitrosylationmentioning

confidence: 99%

Nitric Oxide and Cancer Development

et al. 2007

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Abstract:The role of nitric oxide (NO) in cancer development has become a very active research area. This review presents an overview of many studies that implicate an important role of NO in the development of cancer. These include results in experimental models as well as many observations made on NO and inducible nitric oxide synthase (iNOS) in human cancers. Our survey of the literature has allowed us to conclude that in general large levels of NO will kill cancer cells but paradoxically at intermediate to lower levels NO prevents cancer cell apoptosis and enhances tumor growth. This basic tenet has been demonstrated in many experimental models. The mechanistic basis of how intermediate levels of NO mediate tumor development is an active area of research and is the subject of this review. NO mediated S-nitrosylation of key enzymes and regulatory proteins plays a critical role. Key proteins S-nitrosylated which enhance tumor development include several caspases involved in apoptosis, PTEN the tumor suppressor protein, Bcl-2 the mitochondrial protein which protects from apoptosis, OGG1 the DNA repair protein and methionine adenosyl transferase the liver protein which synthesizes adenosylmethionine. The S-nitrosylation of these proteins enhances DNA mutations, prevents cancer cell apoptosis and enhances oncogenic cell growth. (J Toxicol Pathol 2007; 20: 77-92)

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“…Probe DCG-04 -The epoxide inhibitor JPM-565 has been used as an irreversible inhibitor, with broad reactivity toward cysteine proteases (31,35). Based on the structure of this compound, a derivative was developed to include a biotin affinity tag (DCG-04; Fig.…”

Section: Protease Activity Profiling In Drosophila S2 Cells Using Thementioning

confidence: 99%

“…Our objective was to identify DCG-04-reactive polypeptides in Drosophila phagocytes and to characterize and monitor their activities. Active Site Labeling of Cysteine Proteases in Cell Lysates and Detection by Streptavidin Blotting-JPM-565 and DCG-04 were synthesized and purified as previously described (31,32). Cells were harvested by centrifugation at 4°C, washed in Robb's Drosophila PBS pH 6.8 (52), and cell pellets corresponding to 5 ϫ 10 7 cells were frozen at Ϫ80°C.…”

mentioning

confidence: 99%

Functional Proteomics of the Active Cysteine Protease Content in Drosophila S2 Cells

Kocks

Maehr

Overkleeft

et al. 2003

Molecular & Cellular Proteomics

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The fruit fly genome is characterized by an evolutionary expansion of proteases and immunity-related genes. In order to characterize the proteases that are active in a phagocytic Drosophila model cell line (S2 cells), we have applied a functional proteomics approach that allows simultaneous detection and identification of multiple protease species. DCG-04, a biotinylated, mechanism-based probe that covalently targets mammalian cysteine proteases of the papain family was found to detect Drosophila polypeptides in an activity-dependent manner. Chemical tagging combined with tandem mass spectrometry permitted retrieval and identification of these polypeptides. Among them was thiol-ester motif-containing protein (TEP) 4 which is involved in insect innate immunity and shares structural and functional similarities with the mammalian complement system factor C3 and the panprotease inhibitor alpha2-macroglobulin. We also found four cysteine proteases with homologies to lysosomal cathepsin (CTS) L, K, B, and F, which have been implicated in mammalian adaptive immunity. The Drosophila CTS equivalents were most active at a pH of 4.5. This suggests that Drosophila CTS are, similar to their mammalian counterparts, predominantly active in lysosomal compartments. In support of this concept, we found CTS activity in phagosomes of Drosophila S2 cells. These results underscore the utility of activity profiling to address the functional role of insect proteases in immunity.

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Mechanistic Studies on the Inactivation of Papain by Epoxysuccinyl Inhibitors

Cited by 75 publications

References 62 publications

Murine Cathepsin F Deficiency Causes Neuronal Lipofuscinosis and Late-Onset Neurological Disease

Murine Cathepsin F Deficiency Causes Neuronal Lipofuscinosis and Late-Onset Neurological Disease

Nitric Oxide and Cancer Development

Functional Proteomics of the Active Cysteine Protease Content in Drosophila S2 Cells

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