Inhibition of recombinant dipeptidyl peptidase III by synthetic hemorphin-like peptides

Chiba, T.; Li, Yaohua; Yamane, Toshimi; Ogikubo, Osamu; Fukuoka, Muneyoshi; Arai, Ryohachi; Takahashi, Sho; Ohtsuka, Takanobu; Ohkubo, Iwao; Matsui, Nobuo

doi:10.1016/s0196-9781(03)00119-0

Cited by 40 publications

(41 citation statements)

References 20 publications

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“…An important role of DPP III in the mammalian pain modulatory system is supported by several recent findings: low levels of DPP III activity were detected in the cerebrospinal fluid of individuals suffering from acute pain (17); DPP III exhibits high in vitro affinity toward the important neuropeptides endomorphin-1 and endomorphin-2 (18); and neuroanatomical studies localized rat DPP III in the superficial laminae of the dorsal horn, where enkephalin-synthesizing neurons and high concentrations of endomorphin-2 are found (19,20).…”

mentioning

confidence: 65%

Entropy-driven binding of opioid peptides induces a large domain motion in human dipeptidyl peptidase III

Bezerra

Dobrovetsky

Viertlmayr

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

161

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Opioid peptides are involved in various essential physiological processes, most notably nociception. Dipeptidyl peptidase III (DPP III) is one of the most important enkephalin-degrading enzymes associated with the mammalian pain modulatory system. Here we describe the X-ray structures of human DPP III and its complex with the opioid peptide tynorphin, which rationalize the enzyme's substrate specificity and reveal an exceptionally large domain motion upon ligand binding. Microcalorimetric analyses point at an entropy-dominated process, with the release of water molecules from the binding cleft ("entropy reservoir") as the major thermodynamic driving force. Our results provide the basis for the design of specific inhibitors that enable the elucidation of the exact role of DPP III and the exploration of its potential as a target of pain intervention strategies.isothermal titration calorimetry | metallopeptidase | peptide binding | X-ray crystallography T he endogenous opioid system, composed of opioid peptides and their receptors, modulates a large number of physiological processes, such as endocrine and immune function, gastrointestinal motility, respiration, reward, stress, complex social behavior (e.g., sexual activity), vulnerability to drug addiction, and most notably the procession and transmission of pain stimuli (nociception) (1, 2). Two major types of endogenous opioid peptides are those containing enkephalin sequences at the N terminus (TyrGly-Gly-Phe-Met/Leu) (3) and, more recently identified, endomorphins 1 and 2 (Tyr-Pro-Trp/Phe-Phe-NH 2 ) (4, 5). Knowledge and control over synthesis and degradation pathways of this class of molecules is prerequisite for the development of new therapies that target pertinent physiological processes.Dipeptidyl peptidase III (DPP III), also known as enkephalinase B, is an enkephalin-degrading enzyme that cleaves dipeptides sequentially from the N termini of substrates (6). All DPP IIIs described thus far contain the unique zinc-binding motif HEXXGH characteristic of metallopeptidase family M49 (7). Enzymes from several human and animal tissues, as well as from lower eukaryotes, were purified and biochemically characterized (8, 9). DPP III is largely found as a cytosolic protein, although membrane association in rat brain and Drosophila melanogaster has been described (10, 11). The 3D structure of the yeast ortholog has recently been determined, revealing a unique protein fold with two lobes forming a wide-open substrate-binding cleft (12). The lack of structural information on peptide complexes, however, left the question of substrate specificity largely unanswered.DPP III purified from monkey brain microsomes is strongly inhibited by the neuropeptide spinorphin (Leu-Val-Val-Tyr-ProTrp-Thr), an endogenous factor isolated from bovine spinal cord that also inhibits other enkephalin-degrading enzymes, such as neutral endopeptidase (NEP, neprilysin), aminopeptidase, and angiotensin-converting enzyme (13). Because of a different mode of action compared with morphine, spinorp...

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mentioning

confidence: 65%

Entropy-driven binding of opioid peptides induces a large domain motion in human dipeptidyl peptidase III

Bezerra

Dobrovetsky

Viertlmayr

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

161

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“…However, some pharmacological experiments link DPP III with pain regulation mechanisms because low levels of DPP III activity have been observed in the cerebrospinal fluid of individuals suffering from acute pain (16). Similarly, high concentrations of DPP III found in the superficial laminae of rat spinal cord dorsal horn (17), as well as the high in vitro affinity shown by the human enzyme toward important neuropeptides, such as enkephalins and endomorphins (18), also indicate that this hydrolase could play a role in the mammalian pain modulatory system. These findings make DPP III a potential drug target, and efforts toward inhibitor design and synthesis are under way (19).…”

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confidence: 95%

The First Structure of Dipeptidyl-peptidase III Provides Insight into the Catalytic Mechanism and Mode of Substrate Binding

Baral

Jajčanin-Jozić

Deller

et al. 2008

Journal of Biological Chemistry

111

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Dipeptidyl-peptidases III (DPP III) are zinc-dependent enzymes that specifically cleave the first two amino acids from the N terminus of different length peptides. In mammals, DPP III is associated with important physiological functions and is a potential biomarker for certain types of cancer. Here, we present the 1.95-Å crystal structure of yeast DPP III representing the prototype for the M49 family of metallopeptidases. It shows a novel fold with two domains forming a wide cleft containing the catalytic metal ion. DPP III exhibits no overall similarity to other metallopeptidases, such as thermolysin and neprilysin, but zinc coordination and catalytically important residues are structurally conserved. Substrate recognition is accomplished by a binding site for the N terminus of the peptide at an appropriate distance from the metal center and by a series of conserved arginine residues anchoring the C termini of different length substrates.

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“…Based on the large proteomics datasets measured from seven human cell lines, DPP III is identified as a member of the human central proteome (Burkard et al , 2011 ). It is thought that this peptidase participates in normal intracellular protein catabolism, in the pain modulation, and in endogenous defense against oxidative stress (Chiba et al , 2003 ;Bar š un et al, 2007 ;Liu et al , 2007 ). A pathological role is indicated for DPP III in cataractogenesis and in malignant growth (Zhang et al , 2001 ;Š imaga et al, 2003 ).…”

Section: Introductionmentioning

confidence: 99%

Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents

Karačić¹,

Špoljarić²,

Rožman

et al. 2012

Biological Chemistry

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: Human dipeptidyl peptidase III (DPP III) is a member of the metallopeptidase family M49, involved in protein metabolism and oxidative stress response. DPP III crystal structure shows the two lobe-like domains separated by a wide cleft. The human enzyme has a total of six cysteines, three in the lower (Cys19, Cys147, and Cys176) and three in the upper (Cys509, Cys519, and Cys654), catalytic, domain containing the activesite zinc ion. To elucidate the molecular basis of this enzyme ' s susceptibility to sulfhydryl reagents, biochemical analysis of a set of Cys to Ala mutants was used, supported by mass spectrometry. Cys176, a residue 44 Å apart from the catalytic center of the ligand-free enzyme, was found responsible for the inactivation with the submicromolar concentration of an organomercurial compound, and three additional cysteines contributed to sensitivity to aromatic disulfides. Upon treatment with oxidized glutathione [glutathione disulfide (GSSG)], cysteine residues at positions 147, 176, and 654 were found glutathionylated. The mutational analysis confirmed the involvement of Cys176 and Cys654 in human DPP III inactivation by GSSG. Observation that Cys176, a residue quite distant from the active center, contributes to enzyme inactivation, indicates that the substrate-binding site of human DPP III comprises both lower and upper protein domain.

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Inhibition of recombinant dipeptidyl peptidase III by synthetic hemorphin-like peptides

Cited by 40 publications

References 20 publications

Entropy-driven binding of opioid peptides induces a large domain motion in human dipeptidyl peptidase III

Entropy-driven binding of opioid peptides induces a large domain motion in human dipeptidyl peptidase III

The First Structure of Dipeptidyl-peptidase III Provides Insight into the Catalytic Mechanism and Mode of Substrate Binding

Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents

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