Isolation and characterization of an enkephalin-degrading aminopeptidase from rat brain

Schnebli, Hans Peter; Phillipps, Mildred A.; Barclay, Ralph K.

doi:10.1016/0005-2744(79)90084-6

Cited by 88 publications

(43 citation statements)

References 18 publications

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“…Following inactivation by metal ion chelation, the activity of PAM-1 could be recovered using Zn 2ϩ , Ni 2ϩ , and Co 2ϩ . Both Zn 2ϩ and Co 2ϩ were able to restore activity to metal ion-depleted mammalian PSA (57). Physiochemical analysis would be required to determine which of these metal ions reside within the active site, although the profile of recovery observed with PAM-1 would suggest Zn 2ϩ as the most likely candidate.…”

Section: Fig 3 Silencing Pam-1 By Rnai Shows That It Has Roles In Rmentioning

confidence: 99%

The Caenorhabditis elegans Orthologue of Mammalian Puromycin-sensitive Aminopeptidase Has Roles in Embryogenesis and Reproduction

Brooks

Hooper

Isaac

2003

Journal of Biological Chemistry

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Peptide hydrolysis was inhibited by the metal-chelating agent 1,10-phenanthroline and by the aminopeptidase inhibitors actinonin, amastatin, and leuhistin. However, the enzyme was ϳ100-fold less sensitive toward puromycin (IC 50 , 135 M) than other PSA homologues. Following inactivation of the enzyme, aminopeptidase activity was recovered with Zn 2؉ , Co 2؉ , and Ni 2؉ . Silencing expression of pam-1 by RNA interference resulted in 30% embryonic lethality. Surviving adult hermaphrodites deposited large numbers of oocytes throughout the selffertile period. The overall brood size was, however, unaffected. We conclude that pam-1 encodes an aminopeptidase that clusters phylogenetically with the PSAs, despite attenuated sensitivity toward puromycin, and that it functions in embryo development and reproduction of the nematode.

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Section: Fig 3 Silencing Pam-1 By Rnai Shows That It Has Roles In Rmentioning

confidence: 99%

The Caenorhabditis elegans Orthologue of Mammalian Puromycin-sensitive Aminopeptidase Has Roles in Embryogenesis and Reproduction

Brooks

Hooper

Isaac

2003

Journal of Biological Chemistry

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“…The broad tissue distribution of APP-S and other neutral aminopeptidases, as well as their homology and expression in a variety of species (Constam et al, 1995; McLellan et al, 1988; Schulz et al, 2001; Tobler et al, 1997) can be advantageous to ensure complete and rapid prodrug activation, as was previously noted for Val-Ser-cHPMPC in situ (Eriksson et al, 2008). Additionally, APP-S has been shown to have a broad substrate specificity with preference for hydrophobic and basic amino acids, (Hui et al, 1983; Johnson and Hersh, 1990; Schnebli et al, 1979; Sengupta et al, 2006; Wagner et al, 1981) making it an attractive target for future prodrug design.…”

Section: Discussionmentioning

confidence: 99%

Puromycin-sensitive aminopeptidase: An antiviral prodrug activating enzyme

et al. 2010

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Cidofovir (HPMPC) is a broad-spectrum antiviral agent, currently used to treat AIDS-related human cytomegalovirus retinitis. Cidofovir has recognized therapeutic potential for orthopox virus infections, although its use is hampered by its inherent low oral bioavailability. Val-Ser-cyclic HPMPC (Val-Ser-cHPMPC) is a promising peptide prodrug which has previously been shown by us to improve the permeability and bioavailability of the parent compound in rodent models (Eriksson et al. Molecular Pharmaceutics, 2008 vol 5 598-609). Puromycin-sensitive aminopeptidase was partially purified from Caco-2 cell homogenates and identified as a prodrug activating enzyme for Val-Ser-cHPMPC. The prodrug activation process initially involves an enzymatic step where the LValine residue is removed by puromycin-sensitive aminopeptidase, a step that is bestatin-sensitive. Subsequent chemical hydrolysis results in the generation of cHPMPC. A recombinant puromycinsensitive aminopeptidase was generated and its substrate specificity investigated. The k cat for ValpNA was significantly lower than that for Ala-pNA, suggesting that some amino acids are preferred over others. Furthermore, the three-fold higher k cat for Val-Ser-cHPMPC as compared to Val-pNA suggests that the leaving group may play an important role in determining hydrolytic activity. In addition to its ability to hydrolyze a variety of substrates, these observations strongly suggest that puromycin-sensitive aminopeptidase is an important enzyme for activating Val-Ser-cHPMPC in vivo. Taken together, our data suggest that puromycin-sensitive aminopeptidase makes an attractive target for future prodrug design.

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“…10 The purified enzyme hydrolyses the amino terminal tyrosine residue of metenkephalin. 10,11 It is important to note that AP have been usually described as non-specific enzymes that are capable of hydrolysing a broad spectrum of endogenous peptide substrates and arylamide derivatives to different degrees. However, our results, using whole cells without homogenization, demonstrate different patterns of AP activity in response to the addition of oleic and linoleic fatty acids and cholesterol to the culture medium.…”

Section: Discussionmentioning

confidence: 99%

“…The amount of -naphthylamine released as the result of the enzyme activity was coupled with the GBC salt to form a red complex which can be spectrophotometrically determined at 550 nm. 11 The proteins were quantified in triplicate by the method of Bradford, 12 using BSA as a standard. Specific aminopeptidase activities were expressed as nmol of AlaNNap, ArgNNap, CysNNap, LeuNNap, TyrNNap and pGluNNap hydrolysed per min per mg of protein, using a standard curve prepared withnaphthylamine under the corresponding assay conditions.…”

Section: Methodsmentioning

confidence: 99%

Effects of exogenous fatty acids and cholesterol on aminopeptidase activities in rat astroglia

Ramírez‐Expósito

García

Mayas

et al. 2002

Cell Biochemistry & Function

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Several studies have addressed the interaction between fatty acids and lipids with central nervous system peptides. Because aminopeptidases (AP) are involved in the regulation of neuropeptides, this work studies several AP expressed in cultured astroglia, after exogenous addition of oleic and linoleic fatty acids and cholesterol to the culture medium. Alanyl-AP, arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities were analysed in whole cells using the corresponding aminoacyl-beta-naphthylamides as substrates. Oleic acid inhibits alanyl-AP, cystyl-AP and leucyl-AP activities, whereas linoleic acid inhibits alanyl-AP, arginyl-AP and tyrosyl-AP activities. Neither oleic acid nor linoleic acid modifies pyroglutamyl-AP activity. In contrast, cholesterol increases arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities, although it does not modify alanyl-AP activity. The changes reported here suggest that oleic and linoleic fatty acids and cholesterol can modulate peptide activities via their degradation route involving aminopeptidases; each of them being differentially regulated.

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Isolation and characterization of an enkephalin-degrading aminopeptidase from rat brain

Cited by 88 publications

References 18 publications

The Caenorhabditis elegans Orthologue of Mammalian Puromycin-sensitive Aminopeptidase Has Roles in Embryogenesis and Reproduction

The Caenorhabditis elegans Orthologue of Mammalian Puromycin-sensitive Aminopeptidase Has Roles in Embryogenesis and Reproduction

Puromycin-sensitive aminopeptidase: An antiviral prodrug activating enzyme

Effects of exogenous fatty acids and cholesterol on aminopeptidase activities in rat astroglia

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