Characterization of dynorphin A-converting enzyme in human spinal cord. An endoprotease related to a distinct conversion pathway for the opioid heptadecapeptide?

Silberring, Jerzy; Castello, Maria; Nyberg, Fred

doi:10.1016/s0021-9258(19)36612-8

Cited by 55 publications

(5 citation statements)

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“…The neuroblastoma enzyme containing an active cysteine residue was not capable of cleaving a-neoendorphin and showed preference for full size dynorphin A over its (1 -13) fragment (Satoh et al, 1989). The bovine spinal cord dibasic specific peptidase is not inhibited by EDTA up to 1 mM concentration (Silberring & Nyberg, 1989), and neither is the human spinal cord enzyme that also showed preference for dynorphin A over dynorphin B (Silberring et al, 1992a).…”

Section: Discussionmentioning

confidence: 93%

“…in yeast (Mizuno & Matsuo, 1984), rat testes (Chesneau et al, 1994b), human (Nyberg & Silberring, 1990; Silberring et al, 1992a) and bovine spinal cords (Silberring & Nyberg, 1989), human choroid plexus (Nyberg et al, 1991), cerebrospinal fluid (Nyberg et al, 1985), and neuroblastoma cells (Satoh et al, 1989). These enzymes, which are collectively referred to as dynorphin-converting enzymes, have widely different characteristics: among them are cysteine, serine, and metallopeptidases, with pH optima ranging from 5 to 8.8.…”

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confidence: 99%

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Purification and Characterization of a Secreted Arginine-Specific Dibasic Cleaving Enzyme from EL-4 Cells

Csuhai¹,

Safavi²,

Hersh³

1995

Biochemistry

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A secreted dibasic cleaving peptidase capable of converting dynorphins into Leu-enkephalin-Arg6 was purified from the medium of EL-4 mouse thymoma cells. The enzyme is a novel metalloendopeptidase with a neutral pH optimum (6.9) and a molecular weight of approximately 130 000. The dibasic cleaving enzyme was completely inhibited in the presence of 20-50 mM amine buffers, 0.1 mM EDTA, 0.5 mM 1,10-phenanthroline, 0.5 mM N-ethylmaleimide, and 1mM DTNB. Unlike the Kex2 family of proteases, Ca2+ did not activate the endopeptidase, but high concentrations (1 mM) of metal ions such as Cu2+, Ni2+, Zn2+, and Co2+ completely inhibited the enzyme. Inhibition was not seen with 0.2 mM TLCK, 1 mM DTT, and 1 mM PMSF. The enzyme will cleave Arg-Arg and Arg-Lys bonds, but not Lys-Arg or Lys-Lys bonds in identical environments, and no aminopeptidase or carboxypeptidase activity was seen. The size of the substrate does not seem to be a determining factor, since dynorphin A(1-12) is cleaved at a rate similar to prodynorphin B(228-256) containing 29 amino acids. The identity of the residues on either side of the cleavage site influences the rate of processing, as noted by different rates of cleavage for the same size peptides dynorphin A(1-13) vs dynorphin A(1-9) vs beta-neoendorphin. The presence of proline in the P3' (alpha-neoendorphin), P4' (dynorphin A(1-11)), or P5' (bovine adrenal medulla dodecapeptide) position does not prevent cleavage, but neurotensin and its (1-11) fragment containing both P2 and P2' proline residues are not cleaved.

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Section: Discussionmentioning

confidence: 93%

mentioning

confidence: 99%

Purification and Characterization of a Secreted Arginine-Specific Dibasic Cleaving Enzyme from EL-4 Cells

Csuhai¹,

Safavi²,

Hersh³

1995

Biochemistry

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“…a Protein concentrations were measured by using Coomassie Plus Protein Assay Reagent (Pierce) with BSA as a standard. b Enzyme activity was assayed with bovine adrenal medulla peptide (1)(2)(3)(4)(5)(6)(7)(8) as a substrate at 60 µM final concentration in 20 mM potassium phosphate buffer, pH 7.0, by incubating 5-10 µL of the homogenate or appropriate column fractions for 10-15 min at 37 °C. The reaction rate was calculated based on the appearance of the product, Met-enkephalin-Arg 6 , as determined by HPLC.…”

Section: Resultsmentioning

confidence: 99%

“…A group of enzymes believed to be involved in the processing of dynorphin A and/or dynorphin B to generate Leu-enkephalin-Arg 6 were classified as dynorphin converting enzymes (7,8). We previously reported the purification and characterization of a secreted arginine-specific dibasic cleaving enzyme with dynorphin converting enzyme activity from the EL-4 mouse lymphoma cell line (9).…”

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confidence: 99%

Regulation ofN-Arginine Dibasic Convertase Activity by Amines: Putative Role of a Novel Acidic Domain as an Amine Binding Site

1998

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Peptide sequence analysis and cDNA cloning indicate that a previously described mouse arginine-specific dibasic cleaving enzyme (dynorphin converting enzyme) [Csuhai et al. (1995) Biochemistry 34, 12411] is the homologue of N-arginine dibasic convertase (NRDc) isolated from rat testis [Chesneau et al. (1994) J. Biol. Chem. 269, 2056]. A mouse NRDc cDNA exhibited 98% amino acid identity with the rat cDNA. However, within a 74 residue acidic stretch, this identity drops to 82%. Likewise, the corresponding acidic stretch of human NRDc is only 73% identical with that of rat NRDc. To reconcile previously observed kinetic differences between rat and mouse NRDc, the hydrolysis of peptide substrates by the rat, human, and mouse enzymes was compared using phosphate and Tris as buffers. Although the three NRDc's behaved similarly, Tris had a pronounced effect on the kinetics of peptide hydrolysis. With BAM-8, alpha-neoendorphin, and dynorphin B as substrates, Tris increased KM up to 40-fold with little change in Vmax, while with dynorphin A or somatostatin 28 as substrate, Tris caused a decrease in KM of up to 100 fold, again with only a modest change in Vmax. Other amines, including the polyamines putrescine, spermidine, and spermine, all affected NRD convertase activity. It is proposed that amines bind to the acidic stretch found in NRDc, and that quantitative differences in the sensitivity to amines between the rat, mouse, and human enzymes can be at least partially accounted for by differences in their acidic stretch. The role of polyamines as physiological modulators of N-arginine dibasic convertase is considered.

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“…Androgen-induced modulations in dynorphin may be explained, in part, by androgen-induced modulations in dynorphin-converting enzyme (DCE). DCE transforms dynorphins into enkephalins via cleavage of the dynorphin peptide (Silberring et al, 1992), and DCE concentrations are under regulatory control by androgens. For example, chronic nandrolone administration significantly decreases DCE concentrations in regions that typically exhibit an increase in dynorphin concentrations following androgen treatment, including the caudate putamen, hypothalamus and PAG (Magnusson et al, 2007).…”

Section: Dynorphinmentioning

confidence: 99%

Sex Differences in Opioid-Mediated Effects: Role of Androgens

Sharp

Pearson

Smith

2020

Preprint

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An abundance of data indicate there are sex differences in endogenous opioid peptides and opioid receptors, leading to functional differences in sensitivity to opioid-mediated behaviors between males and females. Many of these sex differences are mediated by the effects of gonadal hormones on the endogenous opioid system. Whereas much research has examined the role of ovarian hormones on opioid-mediated endpoints, comparatively less research has examined the role of androgens. This review describes what is currently known regarding the influence of androgens on opioid-mediated endpoints and how androgens may contribute to sex differences in opioid-mediated effects. The review also addresses the clinical implications of androgenic modulation of opioid-mediated behaviors and suggests future lines of research for preclinical and clinical investigators, We conclude that further investigation into androgenic modulation of opioid-mediated effects may lead to new options for addressing conditions such as chronic pain and substance use disorders.

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Characterization of dynorphin A-converting enzyme in human spinal cord. An endoprotease related to a distinct conversion pathway for the opioid heptadecapeptide?

Cited by 55 publications

References 23 publications

Purification and Characterization of a Secreted Arginine-Specific Dibasic Cleaving Enzyme from EL-4 Cells

Purification and Characterization of a Secreted Arginine-Specific Dibasic Cleaving Enzyme from EL-4 Cells

Regulation ofN-Arginine Dibasic Convertase Activity by Amines: Putative Role of a Novel Acidic Domain as an Amine Binding Site

Sex Differences in Opioid-Mediated Effects: Role of Androgens

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