Nanomolar Inhibitors of the Peptidyl Prolyl Cis/Trans Isomerase Pin1 from Combinatorial Peptide Libraries

Wildemann, Dirk; Erdmann, Frank; Alvarez, Birte Hernandez; Stoller, Gerlind; Zhou, Xiao Zhen; Fanghänel, Jörg; Schutkowski, Mike; Lu, Kun Ping; Fischer, Gunter

doi:10.1021/jm060036n

Cited by 82 publications

(95 citation statements)

References 20 publications

(54 reference statements)

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“…To rule out a nonspecific effect of juglone, we used a more specific Pin1 inhibitor consisting of a competitive peptide containing a Pin1-binding sequence able to inhibit Pin1 both in vitro and in intact cells. 31 This PPIn also abrogated TNF-␣-induced priming of ROS production in response to fMLF ( Figure 2C-D), whereas its effect on fMLF stimulation alone was not significant. Importantly, neither juglone nor PPIn inhibited neutrophil ROS production triggered by PMA, a PKC direct activator ( Figure 2E-F), suggesting that these agents do not inhibit NADPH oxidase activity or the PKC-dependent activation pathway, and that they do not scavenge ROS.…”

Section: Pin1 Inhibitors Inhibit Tnf-␣-induced Priming Of Ros Productmentioning

confidence: 86%

“…To raise antibodies against phosphoSer315, phospho-Ser320, and phospho-Ser328, rabbits were injected with the ovalbumin-crosslinked phosphopeptide sequences of these serines by PolyPeptide Laboratories. The Pin1-peptide inhibitor sequence (Ac-Phe-D-Thr(PO3H2)-Pip-Nal-Gln-NH2) 31 was synthesized by PolyPeptide Laboratories.…”

Section: Reagents and Antibodiesmentioning

confidence: 99%

“…32,33 Measurement of ROS production by luminol-amplified chemiluminescence Neutrophils (5 ϫ 10 5 ) were suspended in 0.5 mL of Hanks balanced salt solution containing 10M luminol at 37°C with or without 250nM juglone or Pin1-peptide inhibitor (PPIn; 50M) 31 for 30 minutes. TNF-␣ (20 ng/mL) was added for 20 minutes; then the cells were stimulated with 10 Ϫ7 M fMLF.…”

Section: Neutrophil Fractionationmentioning

confidence: 99%

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The prolyl isomerase Pin1 acts as a novel molecular switch for TNF-α–induced priming of the NADPH oxidase in human neutrophils

Boussetta

Gougerot-Pocidalo

Hayem

et al. 2010

Blood

114

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Neutrophils play a key role in host defense by releasing reactive oxygen species (ROS). However, excessive ROS production by neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase can damage bystander tissues, thereby contributing to inflammatory diseases. Tumor necrosis factor-␣ (TNF-␣), a major mediator of inflammation, does not activate NADPH oxidase but induces a state of hyperresponsiveness to subsequent stimuli, an action known as priming. The molecular mechanisms by which TNF-␣ primes the NADPH oxidase are unknown. Here we show that Pin1, a unique cis-trans prolyl isomerase, is a previously unrecognized regulator of TNF-␣-induced NADPH oxidase hyperactivation. We first showed that Pin1 is expressed in neutrophil cytosol and that its activity is markedly enhanced by TNF-␣. Inhibition of Pin1 activity with juglone or with a specific peptide inhibitor abrogated TNF-␣-induced priming of neutrophil ROS production induced by N-formylmethionyl-leucyl-phenylalanine peptide (fMLF). TNF-␣ enhanced fMLF-induced IntroductionNeutrophils play an important role in host defense against invading pathogens and in inflammation. In response to stimulating agents, such as the bacterial peptide N-formyl-methionyl-leucyl-phenylalanine (fMLF), neutrophils release large amounts of superoxide anions and other reactive oxygen species (ROS) in a phenomenon called the respiratory burst. ROS produced by the neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase play a key role in host defenses, 1-3 but excessive ROS production can damage healthy bystander tissues, thereby contributing to inflammatory diseases, such as rheumatoid arthritis, inflammatory bowel diseases, and acute respiratory distress syndrome. 4,5 Neutrophil ROS production is mediated by the phagocyte NADPH oxidase, also called NOX2. NADPH oxidase is a multicomponent enzyme system that catalyzes NADPH-dependent reduction of oxygen to superoxide anion. 6,7 In resting cells, the NADPH oxidase is inactive and its components are distributed between the cytosol and membranes. When cells are activated, the cytosolic components (p47phox, p67phox, p40phox, and Rac2) migrate to the membranes, where they associate with the membranebound components (p22phox and gp91phox/NOX2, which form the flavocytochrome b558) to assemble the catalytically active oxidase. 7,8 During NADPH oxidase activation, p47phox, p67phox, p40phox, p22phox, and gp91phox/NOX2 become phosphorylated. 9-13 p47phox phosphorylation on several serines plays a pivotal role in oxidase activation in intact cells. 14,15 Neutrophil ROS production is enhanced or primed by a variety of mediators, including proinflammatory cytokines, such as tumor necrosis factor-␣ (TNF-␣). In vitro, TNF-␣ induces a very weak oxidative response by neutrophils but strongly enhances ROS release on exposure to a secondary stimulus, such as the bacterial peptide fMLF. [16][17][18] This "priming" of neutrophil ROS production plays a detrimental role in a variety of human inflammatory diseases, in which ROS hyperp...

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Section: Pin1 Inhibitors Inhibit Tnf-␣-induced Priming Of Ros Productmentioning

confidence: 86%

Section: Reagents and Antibodiesmentioning

confidence: 99%

Section: Neutrophil Fractionationmentioning

confidence: 99%

See 1 more Smart Citation

The prolyl isomerase Pin1 acts as a novel molecular switch for TNF-α–induced priming of the NADPH oxidase in human neutrophils

Boussetta

Gougerot-Pocidalo

Hayem

et al. 2010

Blood

114

View full text Add to dashboard Cite

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“…More specific Pin1 inhibitors with less side effects are required. Promising candidates are the substituted aryl 1-indanyl ketones (53) and a series of specific D-phospho-Thr-containing peptides (54,55). Such inhibitors might not only be useful in studying the functional relevance of Pin1 in vivo; they could also provide the therapeutic basis to treat pathological conditions in which Pin1 is aberrantly upregulated, such as cancer (8).…”

Section: Discussionmentioning

confidence: 99%

Juglone Inactivates Cysteine-rich Proteins Required for Progression through Mitosis

Fila

Metz

Sluijs

2008

Journal of Biological Chemistry

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The parvulin peptidyl-prolyl isomerase Pin1 catalyzes cistrans isomerization of p(S/T)-P bonds and might alter conformation and function of client proteins. Since the trans conformation of p(S/T)-P bonds is preferred by protein phosphatase 2A (PP2A), Pin1 may facilitate PP2A-mediated dephosphorylation. Juglone irreversibly inhibits parvulins and is often used to study the function of Pin1 in vivo. The drug prevents dephosphorylation of mitotic phosphoproteins, perhaps because they bind Pin1 and are dephosphorylated by PP2A. We show here however that juglone inhibited post-mitotic dephosphorylation and the exit of mitosis, independent of Pin1. This effect involved covalent modification of sulfhydryl groups in proteins essential for metaphase/anaphase transition. Particularly cytoplasmic proteins with a high cysteine content were vulnerable to the drug. Alkylation of sulfhydryl groups altered the conformation of such proteins, as evidenced by the disappearance of antibody epitopes on tubulin and the mitotic checkpoint component BubR1. The latter activates the anaphase-promoting complex/ cyclosome, which degrades regulatory proteins, such as cyclin B1 and securins, and is required for mitotic exit. Indeed, juglone-treated cells failed to assemble a mitotic spindle, which correlated with perturbed microtubule dynamics, loss of immunodetectable tubulin, and formation of tubulin aggregates. Juglone also prevented degradation of cyclin B1, independently of the Mps1-controlled mitotic spindle checkpoint. Since juglone affected cell cycle progression at several levels, more specific drugs need to be developed for studies of Pin1 function in vivo. Peptidyl-prolyl isomerases (PPIases)2 accelerate the cis-trans conversion of peptide bonds preceding prolyl residues, which can cause alterations in protein conformation (e.g. see Refs. 1 and 2). PPIases have been grouped into cyclophilin, FK506-binding protein, and parvulin subfamilies (see Ref. 3), for which distinct pharmacological inhibitors are available. The parvulin group is irreversibly inhibited by juglone (4). Pin1 comprises an N-terminal type IV WW domain, which determines phosphorylation-specific protein-protein interactions, and a C-terminal PPIase domain that harbors the catalytic center (see Ref. 5). Pin1 is a unique PPIase, because it preferably binds to side chain-phosphorylated S/T-P moieties in numerous proteins, including crucial cell cycle regulators or proteins that become phosphorylated immediately prior to cell division (6, 7). Isomerization of the p(S/T)-P peptide bond regulates, for instance, localization and phosphorylation status of Pin1 client proteins (see Ref. 5). Pin1 is therefore a regulator that, in concert with proline-directed kinases, phosphatases, and ubiquitin ligases, controls the cell cycle (see Refs. 5 and 8). Pin1 is possibly a cancer target gene, because its overexpression enhances transformed phenotypes induced by oncogenic Ras and Neu (see (5)). Pin1 is overexpressed in many human cancers, and its overexpression correlates with poor...

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“…A few years later, a series of potent phosphorylated pentameric to octameric phosphorylated peptidic inhibitors of Pin1 activity was selected from a combinatorial library (Wildemann et al, 2006a). These inhibitors contained non-proteogenic amino acids and blocked cell cycle progression in HeLa cells in a dose-dependent manner.…”

Section: Peptidic Inhibitors and Substrate Analogsmentioning

confidence: 99%

Structure and function of the human parvulins Pin1 and Par14/17

Matena

Rehic

Hönig

et al. 2018

Biological Chemistry

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Parvulins belong to the family of peptidyl-prolyl cis/trans isomerases (PPIases) assisting in protein folding and in regulating the function of a broad variety of proteins in all branches of life. The human representatives Pin1 and Par14/17 are directly involved in processes influencing cellular maintenance and cell fate decisions such as cell-cycle progression, metabolic pathways and ribosome biogenesis. This review on human parvulins summarizes the current knowledge of these enzymes and intends to oppose the well-studied Pin1 to its less wellexamined homolog human Par14/17 with respect to structure, catalytic and cellular function.

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Nanomolar Inhibitors of the Peptidyl Prolyl Cis/Trans Isomerase Pin1 from Combinatorial Peptide Libraries

Cited by 82 publications

References 20 publications

The prolyl isomerase Pin1 acts as a novel molecular switch for TNF-α–induced priming of the NADPH oxidase in human neutrophils

The prolyl isomerase Pin1 acts as a novel molecular switch for TNF-α–induced priming of the NADPH oxidase in human neutrophils

Juglone Inactivates Cysteine-rich Proteins Required for Progression through Mitosis

Structure and function of the human parvulins Pin1 and Par14/17

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