Substrate specificity of the protease that processes human interleukin-1ß

Sleath, Paul R.; Hendrickson, Ronald C.; Kronheim, С.; March, Carl J.; Black, Roy A.

doi:10.1007/978-94-011-3034-9_330

Cited by 44 publications

(52 citation statements)

References 3 publications

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“…VEMD/VEVD3 VERD in K14, K18, or K19) or A3 D mutations (VEMDA/VEVDA3 VEMDD/VEVDD in K14 or K18), which mimic K14 mutations found in EBS patients, had no measurable effect on caspase-mediated cleavage of the type I keratin. These results, using an in vivo transfection system, are similar to what has been noted with in vitro peptide substrates in that the X 2 position can tolerate a wide range of amino acids, whereas X 1 seems to dictate caspase enzyme-type specificity (27)(28)(29). In contrast, the X 3 position tolerated an A3 D substitution well, which would not have been predicted based on peptide in vitro substrate studies (28,29).…”

Section: Apoptosis-associated Degradation Of Keratins-k18 (5-7)supporting

confidence: 81%

Effect of Mutation and Phosphorylation of Type I Keratins on Their Caspase-mediated Degradation

Omary

2001

Journal of Biological Chemistry

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Type I keratins K18 and K19 undergo caspase-mediated degradation during apoptosis. Two known K18 caspase cleavage sites are aspartates in the consensus sequences VEVDA and DALDS, located within the rod domain and tail domain, respectively. Several K14 (another type I keratin) mutations within the caspase cleavage motif have been described in patients with epidermolysis bullosa simplex. Here we use extensive mutational analysis to show that K19 and K14 are caspase substrates and that the ability to undergo caspase-mediated digestion of K18, K19, or K14 is highly dependent on the location and nature of the mutation within the caspase cleavage motif. proteins of epithelial cells and consist of Ͼ20 separate gene products (1). The keratin subfamily of IF proteins is classified into two major groups, type I keratins (K9 -20) and type II keratins (K1-8), which associate as noncovalent type I-II heteropolymers in an epithelial cell type-specific manner (1-4). Among cytoplasmic IF proteins, keratins and vimentin undergo caspase-mediated degradation as part of the cytoskeletal remodeling that takes place during apoptosis (5-8). The nuclear lamin IF proteins also undergo degradation during apoptosis and were the first IF proteins demonstrated to undergo apoptosis-associated digestion (9 -11). The only keratins shown to undergo proteolysis during apoptosis are K18 and K19, whereas their type II partner (i.e. K8) manifests remarkable resistance to apoptotic degradation. Two known K18 caspase sites, VEVD and DALD, are located in the rod domain and tail domain, respectively (5-7). VEVD or similar consensus sequences are found in other IF proteins within the so-called linker 1-2 (L1-2) region of the rod domain (Fig. 1), whereas DALD is a unique caspase site that is found only in the K18 tail domain. The signals, if any, that target keratin degradation and the significance of this proteolysis are unknown. To that end, the only keratin-related apoptosis-associated change after an apoptotic signal is marked early keratin hyperphosphorylation. The significance of this early keratin hyperphosphorylation in association with apoptosis is not known, but amino acid substitution of the major K18 phosphorylation sites does not alter susceptibility to caspase digestion (6).Understanding the significance and regulation of keratin (and other IF protein) degradation during apoptosis is important from a cell biological perspective and may also have pathophysiological relevance to human disease. For example, although most keratin mutations that have been identified in patients with epidermal blistering keratin diseases are located at the N-terminal region of the rod IA subdomain (12, 13), at least four K14 mutations have been described within the L1-2 region (14 -17) in close proximity to the caspase recognition motif (VEMDA, also referred to herein as the caspase box). The cause of blister formation in these patients may be attributed to keratin filament assembly defects with resultant cell fragility. However, the proximity of these mutations ...

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Section: Apoptosis-associated Degradation Of Keratins-k18 (5-7)supporting

confidence: 81%

Effect of Mutation and Phosphorylation of Type I Keratins on Their Caspase-mediated Degradation

Omary

2001

Journal of Biological Chemistry

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“…Inactive precursors of ICE-related proteases are cleaved at aspartate residues to become active, resulting in apoptosis. [39][40][41] We are pursuing the possibility that ICE-related protein cleavage characterizes lovastatin-induced apoptosis of medulloblastoma cells. 32 Interfering directly with the function of ICE-related proteases or their precursors might inhibit cell death, and it is conceivable that mevalonate inhibition of lovastatin-induced apoptosis is simply the result of such a phenomenon.…”

Section: Discussionmentioning

confidence: 99%

Mevalonate Prevents Lovastatin-Induced Apoptosis in Medulloblastoma Cell Lines

Wang

Macaulay²

1999

Can. j. neurol. sci.

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Background: 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) is a key rate-limiting enzyme in the mevalonate pathway, which generates precursors for cholesterol biosynthesis and the production of non-steroidal mevalonate derivatives that are involved in a number of growth-regulatory processes. We have reported that lovastatin, a competitive inhibitor of HMG-CoA reductase, not only inhibits medulloblastoma proliferation in vitro, but also induces near-complete cell death via apoptosis. The present study explores some of the pathways which may be involved in lovastatin-induced apoptosis. Methods: Medulloblastoma cell lines were exposed in vitro to lovastatin with or without mevalonate, and document the effects using morphology, flow cytometry, DNA electrophoresis and Northern analysis. Results: 1) Mevalonate prevents apoptosis when co-incubated with lovastatin, or when administered to lovastatin-pretreated cells. 2) Mevalonate restores the lovastatinarrested cell cycle, allowing S phase entry. 3) Mevalonate does not prevent lovastatin-induced apoptosis after a critical duration of lovastatin pretreatment. For cell lines Daoy and UW228 this was 24 hours, and for D283 Med and D341 Med it was 48 hours. 4) Increases in HMG-CoA reductase mRNA levels induced by lovastatin are abrogated by co-incubation with lovastatin and mevalonate. Conclusions: These results confirm that lovastatin inhibition of this enzyme results in blockage of the mevalonate pathway, and that such a block is a critical step in the mechanism of lovastatin-induced apoptosis.RÉSUMÉ: Le mévalonate prévient l'apoptose induite par la lovastatine dans des lignées cellulaires de médulloblastome. Introduction: La 3-hydroxy-3-méthylglutaryl coenzyme a réductase (HMG-CoA réductase) est une enzyme clé, qui a un rôle d'étape cinétiquement limitante dans la voie du mévalonate et qui génère des précurseurs pour la biosynthèse du cholestérol et la production de dérivés non stéroïdiens du mévalonate impliqués dans certains processus de régulation de la croissance. Nous avons rapporté que la lovastatine, un inhibiteur compétitif de l'HMG-CoA réductase, non seulement inhibe la prolifération du médulloblastome in vitro, mais également induit presque complètement la mort cellulaire via l'apoptose. Cette étude explore certaines des voies qui pourraient être impliquées dans l'apoptose induite par la lovastatine. Méthodes: Les effets de l'exposition in vitro de lignées de cellules de médulloblastome à la lovastatine avec ou sans mévalonate ont été évalués par des études morphologiques, par cytométrie de flux, électrophorèse de l'ADN et analyse de Northern. Résultats: 1) Le mévalonate prévient l'apoptose quand les cellules sont coincubées avec la lovastatine ou quand il est administré à des cellules prétraitées par la lovastatine. 2) Le mévalonate rétablit le cycle cellulaire interrompu par la lovastatine, permettant une entrée en phase S. 3) Le mévalonate ne prévient pas l'apoptose induite par la lovastatine après un temps de prétraitement cri...

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“…Caspase-1 is a novel type of cysteine protease containing an active-site cysteine residue (Cys-285) in the p20 subunit, the mutation of which results in loss of activity. Substrate specificity studies revealed that caspase-1 has a strong preference for aspartic acid adjacent to the cleavage site in the P " position, a small hydrophobic amino acid (Gly or Ala) in the Ph " position, and also a requirement for four amino acids to the left of the cleavage site [33,39]. AcetylTyr-Val-Ala-Asp-CHO (Ac-YVAD.CHO, where CHO is aldehyde) and Ac-YVAD-7-amino-4-methylcoumarin (Ac-YVAD-AMC) were synthesized as a potent competitive reversible inhibitor and a fluorimetric substrate respectively for caspase-1 [33].…”

Section: Structure and Functionmentioning

confidence: 99%

Caspases: the executioners of apoptosis

Cohen

1997

Biochemical Journal

4,259

2,909

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Apoptosis is a major form of cell death, characterized initially by a series of stereotypic morphological changes. In the nematode Caenorhabditis elegans, the gene ced-3 encodes a protein required for developmental cell death. Since the recognition that CED-3 has sequence identity with the mammalian cysteine protease interleukin-1β-converting enzyme (ICE), a family of at least 10 related cysteine proteases has been identified. These proteins are characterized by almost absolute specificity for aspartic acid in the P " position. All the caspases (ICE-like proteases) contain a conserved QACXG (where X is R, Q or G) pentapeptide activesite motif. Caspases are synthesized as inactive proenzymes comprising an N-terminal peptide (prodomain) together with one large and one small subunit. The crystal structures of both caspase-1 and caspase-3 show that the active enzyme is a heterotetramer, containing two small and two large subunits. Activation of caspases during apoptosis results in the cleavage of critical cellular substrates, including poly(ADP-ribose) poly-

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Substrate specificity of the protease that processes human interleukin-1ß

Cited by 44 publications

References 3 publications

Effect of Mutation and Phosphorylation of Type I Keratins on Their Caspase-mediated Degradation

Effect of Mutation and Phosphorylation of Type I Keratins on Their Caspase-mediated Degradation

Mevalonate Prevents Lovastatin-Induced Apoptosis in Medulloblastoma Cell Lines

Caspases: the executioners of apoptosis

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