Human dipeptidyl peptidase III acts as a post-proline-cleaving enzyme on endomorphins

Baršun, Marina; Jajčanin, Nina; Vukelić, Bojana; Špoljarić, Jasminka; Abramić, Marija

doi:10.1515/bc.2007.039

Cited by 61 publications

(66 citation statements)

References 36 publications

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“…In contrast, all subsites in DPP III are deep and somewhat hydrophobic (SI Appendix, Fig. S7 and Table S2), in agreement with the relaxed specificity shown by this enzyme (18).…”

Section: Resultssupporting

confidence: 77%

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.

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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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“…In contrast, all subsites in DPP III are deep and somewhat hydrophobic (SI Appendix, Fig. S7 and Table S2), in agreement with the relaxed specificity shown by this enzyme (18).…”

Section: Resultssupporting

confidence: 77%

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.

Self Cite

161

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“…3). Although DPP III enzymes are known to be quite unspecific with respect to sequence and length variations of the peptide substrates (18,48,49), they strictly cleave off two amino acid residues from the N terminus of their substrate, i.e. they act in a dipeptidyl-peptidase manner.…”

Section: Discussionmentioning

confidence: 99%

“…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).…”

mentioning

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

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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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“…DPP III was purified and biochemically characterized from several human and animal tissues, slime mold, Drosophila and the yeast Saccharomyces cerevisiae 2 . It was shown that this cytosolic enzyme, which is broadly distributed in eukaryotic cells and tissues, in vitro hydrolyzes a number of biologically active peptides, including opioid peptides enkephalins and endomorphins 1,4,5 . Therefore, its contribution in normal intracellular protein catabolism was assumed and the role in the endogenous pain modulation was suggested.…”

Section: Introductionmentioning

confidence: 99%

Novel dipeptidyl hydroxamic acids that inhibit human and bacterial dipeptidyl peptidase III

Cvitešić¹,

Sabljić

Makarević³

et al. 2016

Journal of Enzyme Inhibition and Medicinal Chemistry

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Human dipeptidyl peptidase III (hDPP III), a zinc-metallopeptidase of the family M49, is an activator of the Keap1-Nrf2 cytoprotective pathway involved in defense against oxidative stress. Pathophysiological roles of DPP III have not been elucidated yet, partly due to the lack of specific inhibitors. We showed that substrate analog H-Tyr-Phe-NHOH is a strong competitive inhibitor of hDPP III, while H-Tyr-Gly-NHOH expresses much weaker inhibition. To investigate the effects of amino acid substitutions in inhibitor P1 position, we synthesized three new dipeptidyl hydroxamates and examined their influence on the activity of hDPP III and DPP III from the human gut symbiont Bacteroides thetaiotaomicron. The extent of inhibition of hDPP III, but not of bacterial enzyme, was dependent on the amino acid in P1. H-Phe-Phe-NHOH is recognized as one of the strongest inhibitors of hDPP III (K i ¼ 0.028 mM), and H-Phe-Leu-NHOH discriminated between human and bacterial ortholog of the M49 family.

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Human dipeptidyl peptidase III acts as a post-proline-cleaving enzyme on endomorphins

Cited by 61 publications

References 36 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

Novel dipeptidyl hydroxamic acids that inhibit human and bacterial dipeptidyl peptidase III

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