N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a natural tetra-peptide with anti-inflammatory and anti-fibrotic properties. We have previously shown that prolyl-oligopeptidase (POP) is involved in the Ac-SDKP release from thymosin β4 (Tβ4). However, POP can only hydrolyze peptides shorter than 30 amino acids and Tβ4 is 43 amino acids long. This indicates that before POP hydrolysis takes place, Tβ4 is hydrolyzed by another peptidase that releases N-terminal intermediate peptide(s) less than 30 amino acids. Our peptidase database search pointed out meprin α metalloprotease as a potential candidate. Therefore, we hypothesized that prior to POP hydrolysis, Tβ4 is hydrolyzed by meprin α. In vitro, we found that the incubation of Tβ4 with both, meprin α and POP released Ac-SDKP, whereas no Ac-SDKP was released when Tβ4 was incubated with either meprin α or POP alone. Incubation of Tβ4 with rat kidney homogenates significantly released Ac-SDKP, which was blocked by meprin α inhibitor, actinonin. In addition, kidneys from meprin α knockout (KO) mice showed significantly lower basal Ac-SDKP amount, compared to wild-type mice. Kidney homogenates from meprin α KO mice failed to release Ac-SDKP from Tβ4. In vivo, we observed that, rats treated with ACE inhibitor, captopril, increased plasma concentrations of Ac-SDKP, which was inhibited by the co-administration of actinonin (vehicle 3.1±0.2 nmol/L; captopril 15.1±0.7 nmol/L; captopril + actinonin 6.1±0.3 nmol/L; P<0.005). Similar results were obtained with urinary Ac-SDKP after actinonin treatment. We conclude that, release of Ac-SDKP from Tβ4 is mediated by successive hydrolysis involving meprin α and POP.
- THYMOSIN BETA 4
- MEPRIN ALPHA
- ANGIOTENSIN CONVERTING ENZYME
- Prolyl oligopeptidase
- Copyright © 2015, American Journal of Physiology - Renal Physiology