| Full text | |
| Author(s): |
Dalio, Fernanda M.
[1]
;
Machado, Mauricio F. M.
[2]
;
Marcondes, Marcelo F.
[1]
;
Juliano, Maria A.
[1]
;
Chagas, Jair R.
[1]
;
Cunha, Rodrigo L. O. R.
[3]
;
Oliveira, Vitor
[1]
Total Authors: 7
|
| Affiliation: | [1] Univ Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo, SP - Brazil
[2] Univ Mogi das Cruzes, CIIB, BR-08780911 Mogi Das Cruzes, SP - Brazil
[3] Univ Fed ABC, CCNH, Lab Biol Quim, BR-09210170 Santo Andre, SP - Brazil
Total Affiliations: 3
|
| Document type: | Journal article |
| Source: | Biochemical and Biophysical Research Communications; v. 522, n. 2, p. 368-373, FEB 5 2020. |
| Web of Science Citations: | 0 |
| Abstract | |
Thimet oligopeptidase (TOP, EC 3.4.24.15) and neurolysin (NEL, EC 3.4.24.16) are closely related zinc-dependent metalo-oligopeptidases, which take part in the metabolism of oligopeptides (from 5 to 17 amino acid residues) inside and outside cells. Both peptidases are ubiquitously distributed in tissues. TOP is one of the main intracellular peptide-processing enzymes being important for the antigen selection in the MHC Class I presentation route, while NEL function has been more associated with the extracellular degradation of neurotensin. Despite efforts being made to develop specific inhibitors for these peptidases, the most used are: CPP-Ala-Ala-Tyr-PABA, described by Orlowski et al. in 1988, and CPP-Ala-Aib-Tyr-PABA (JA-2) that is an analog more resistant to proteolysis, which development was made by Shrimpton et al. in 2000. In the present work, we describe other analogs of these compounds but, with better discriminatory capacity to inhibit specifically NEL or TOP. The modifications introduced in these new analogs were based on a key difference existent in the extended binding sites of NEL and TOP: the negatively charged Glu(469) residue of TOP corresponds to the positively charged Arg(470) residue of NEL. These residues are in position to interact with the residue at the P-1' and/or P-2' of their substrates (mimicked by the Ala-Ala/P1'-P2' residues of the CPP-Ala-Ala-Tyr-PABA). Therefore, exploring this single difference, the following compounds were synthesized: CPP-Asp-Ala-Tyr-PABA, CPP-Arg-Ala-Tyr-PABA, CPP-Ala-Asp-Tyr-PABA, CPP-Ala-Arg-Tyr-PABA. Confirming the predictions, the replacement of each noncharged residue of the internal portion Ala-Ala by a charged residue Asp or Arg resulted in compounds with higher selectivity for NEL or TOP, especially due to the electrostatic attraction or repulsion by the NEL Arg(470) or TOP Glu(469) residue. The CPP-Asp-Ala-Tyr-PABA and CPP-Ala-Asp-Tyr-PABA presented higher affinities for NEL, and, the CFP-Ala-Arg-Tyr-PABA showed higher affinity for TOP. (C) 2019 Elsevier Inc. All rights reserved. (AU) | |
| FAPESP's process: | 19/05936-0 - Production of yeast metacaspases with N-terminal depletion and containing site-directed mutations and biochemical characterization of these enzymes |
| Grantee: | Mauricio Ferreira Marcondes Machado |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 18/09158-0 - Prolyl oligopeptidase: structure-activity and secretion |
| Grantee: | Vitor Marcelo Silveira Bueno Brandão de Oliveira |
| Support Opportunities: | Regular Research Grants |