Structure–activity studies with high-affinity inhibitors of pyroglutamyl-peptidase II

Kelly, Julie; Scalabrino, Gaia; Slator, Gillian R.; Cullen, Aoife A.; Gilmer, John F.; Lloyd, David G.; Bennett, Gerald; Bauer, Karl; Tipton, Keith F.; Williams, C. H.

doi:10.1042/bj20041722

Cited by 10 publications

(11 citation statements)

References 44 publications

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“…Recent results suggest a ligand binding mode for TRH in PPII active site that differs from the orientation that bestatin adopts in the active site of LTA4H structure (44). The binding mode of bestatin is the binding orientation proposed for substrates of M1 aminopeptidases based on theoretical and experimental data (31)(32)(33)36).…”

Section: Methodsmentioning

confidence: 99%

Homology Modeling and Site-directed Mutagenesis of Pyroglutamyl Peptidase II

Chávez‐Gutiérrez¹,

Matta‐Camacho²,

Osuna³

et al. 2006

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Homology Modeling and Site-directed Mutagenesis of Pyroglutamyl Peptidase II

Chávez‐Gutiérrez¹,

Matta‐Camacho²,

Osuna³

et al. 2006

Journal of Biological Chemistry

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“…Since it has been shown that most peptidases bind to their substrate in one direction at their catalytic center (8,9), there could be only one direction for the collagen triple helices at the binding site of CBD. On the other hand, CBD might allow bidirectional binding, since it is independent of the catalytic domain.…”

Section: ؉mentioning

confidence: 99%

Unidirectional Binding of Clostridial Collagenase to Triple Helical Substrates

Philominathan

Koide

Hamada

et al. 2009

Journal of Biological Chemistry

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Histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. The C-terminal collagen-binding domain (CBD) of these enzymes is the minimal segment required to bind to collagen fibril. Collagen binding efficiency of CBD is more pronounced in the presence of Ca Histotoxic clostridia produce collagenases that degrade collagen in connective tissue. Although the enzyme is assumed to be a causative agent for diseases like gas gangrene (1), it is beneficial to remove dead tissue from ulcers or burns and for nonsurgical treatment of Dupuytren's disease (2, 3). For collagenases to hydrolyze tissue collagen, the enzymes must 1) anchor themselves onto an insoluble collagen fibril, which is a staggered array of tropocollagen and then 2) isolate a single triple helical molecule from the bundle and finally 3) unwind the triple helix to expose a scissile peptide bond. Clostridium histolyticum produces two classes of collagenases, which contain a catalytic domain belonging to the family M9B, followed by one or two copies of polycystic kidney disease domains and one or two copies of collagen-binding domains (CBD) 2 (4). Each CBD spans ϳ120 amino acid residues and binds specifically to insoluble collagen. CBD also binds to collagenous peptides with triple helical conformation but not to collagenous peptides that lack triple helix or to gelatin (denatured collagen), suggesting that the CBD-collagen interaction is conformation-specific (4, 5). Calcium ions enhance the binding at physiological concentration, and x-ray crystal structures of CBD have been solved in the presence and absence of calcium (6).Since collagen fibrils constitute a major part of the extracellular matrix, bioactive molecules can be anchored with CBD for their prolonged effect. Nishi et al. (7) have demonstrated that growth factors fused to CBD remained at the sites of injection much longer than growth factors alone to induce extended cell proliferation. In order to gain an insight into the anchoring mechanism of CBD, we attempted to co-crystallize CBD and collagenous peptide without success. Also to better address the role of CBD in fibril disruption and transition states from insoluble substrate, solution studies of CBD with the triple helical collagenous peptide became necessary.NMR titration methods were utilized to identify the collagen binding pocket on CBD. Since it has been shown that most peptidases bind to their substrate in one direction at their catalytic center (8, 9), there could be only one direction for the collagen triple helices at the binding site of CBD. On the other hand, CBD might allow bidirectional binding, since it is independent of the catalytic domain. To identify the binding direction, three different NMR titrations were performed with spinlabeled analogues of tropocollagen, where a nitroxide spin label 2,2,5,5-tetramethyl-L-pyrrolidinyloxy (PROXYL) was attached to either the N or C terminus of the collagenous peptide. The nitroxide moiety with an unpaired electron can cause enhancement in pa...

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“…Neuropeptides, though, are often susceptible to proteolytic degradation, which is a major impediment to their therapeutic use (Vlieghe et al, 2010). In the case of TRH, susceptibility to degradation and potential for endocrine side effects have critically limited its clinical application (Kelly et al, 2000(Kelly et al, , 2005Kelly, 1995; al., 2007). By overcoming the inadequacies of TRH and prior TRH analogs, JAK4D now presents the possibility of realizing the benefits offered by activation of the central TRH signaling system in the treatment of CNS disorders (Scalabrino et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

“…Membranes from these tissues were prepared at the University of Miami and purchased from Tissue Solutions Limited, Scotland. Membranes were prepared essentially as described by (Hirst et al, 2000) except homogenization and resuspension buffers were the same as those we employed previously (Hogan et al, 2008;Kelly et al, 2005Kelly et al, , 2002. Membranes were stored at -80 °C pending assay.…”

Section: Radioligand Binding Studiesmentioning

confidence: 99%

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First-in-class thyrotropin-releasing hormone (TRH)-based compound binds to a pharmacologically distinct TRH receptor subtype in human brain and is effective in neurodegenerative models

Kelly¹,

Boyle²,

Cole³

et al. 2015

Neuropharmacology

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Hardiman, O., First-in-class thyrotropin-releasing hormone (TRH)-based compound binds to a pharmacologically distinct TRH receptor subtype in human brain and is effective in neurodegenerative models, Neuropharmacology (2014), doi: 10.1016/j.neuropharm.2014.09.024. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPT M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPTABSTRACT JAK4D, a first-in-class thyrotropin-releasing hormone (TRH)-based compound, is a prospective therapeutic candidate offering a multifaceted approach to treating neurodegeneration and other CNS conditions. The purpose of these studies was to determine the ability of JAK4D to bind to TRH receptors in human brain and to evaluate its neuropharmacological effects in neurodegenerative animal models. Additionally, JAK4D brain permeation was examined in mouse, and initial toxicology was assessed in vivo and in vitro. We report that JAK4D bound selectively with nanomolar affinity to native TRH receptors in human hippocampal tissue and showed for the first time that these receptors are pharmacologically distinct from TRH receptors in human pituitary, thus revealing a new TRH receptor subtype which represents a promising neurotherapeutic target in human brain. Systemic administration of JAK4D elicited statistically significant and clinically-relevant neuroprotective effects in three established neurodegenerative animal models: JAK4D reduced cognitive deficits when administered post-insult in a kainate (KA)-induced rat model of neurodegeneration; it protected against free radical release and neuronal damage evoked by intrastriatal microdialysis of KA in rat; and it reduced motor decline, weight loss, and lumbar spinal cord neuronal loss in G93A-SOD1 transgenic Amyotrophic Lateral Sclerosis mice. Ability to cross the blood-brain-barrier and a clean initial toxicology profile were also shown. In light of these findings, JAK4D is an important tool for investigating the hitherto-unidentified central TRH receptor subtype reported herein and an attractive therapeutic candidate for neurodegenerative disorders.

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Structure–activity studies with high-affinity inhibitors of pyroglutamyl-peptidase II

Cited by 10 publications

References 44 publications

Homology Modeling and Site-directed Mutagenesis of Pyroglutamyl Peptidase II

Homology Modeling and Site-directed Mutagenesis of Pyroglutamyl Peptidase II

Unidirectional Binding of Clostridial Collagenase to Triple Helical Substrates

First-in-class thyrotropin-releasing hormone (TRH)-based compound binds to a pharmacologically distinct TRH receptor subtype in human brain and is effective in neurodegenerative models

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