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Prolyl oligopeptidase induces angiogenesis both in vitro and in vivo in a novel regulatory manner
Author(s) -
Myöhänen TT,
TenorioLaranga J,
Jokinen B,
VázquezSánchez R,
MorenoBaylach MJ,
GarcíaHorsman JA,
Männistö PT
Publication year - 2011
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/j.1476-5381.2010.01146.x
Subject(s) - oligopeptidase , angiogenesis , in vivo , matrigel , angiogenesis inhibitor , pharmacology , in vitro , chemistry , umbilical vein , proteases , biochemistry , biology , enzyme , cancer research , microbiology and biotechnology
BACKGROUND AND PURPOSE A serine protease, prolyl oligopeptidase (POP) has been reported to be involved in the release of the pro‐angiogenic tetrapeptide acetyl‐N‐Ser‐Asp‐Lys‐Pro (Ac‐SDKP) from its precursor, 43‐mer thymosin β4 (Tβ4). Recently, it was shown that both POP activity and the levels of Ac‐SDKP are increased in malignant tumours. The aim of this study was to clarify the release of Ac‐SDKP, and test if POP and a POP inhibitor, 4‐phenyl‐butanoyl‐L‐prolyl‐2( S )‐cyanopyrrolidine (KYP‐2047), can affect angiogenesis. EXPERIMENTAL APPROACH We used HPLC for bioanalytical and an enzyme immunoassay for pharmacological analysis. Angiogenesis of human umbilical vein endothelial cells was assessed in vitro using a ‘tube formation’ assay and in vivo using a Matrigel plug assay (BD Biosciences, San Jose, CA, USA) in adult male rats. Moreover, co‐localization of POP and blood vessels was studied. KEY RESULTS We showed the sequential hydrolysis of Tβ4: the first‐step hydrolysis by proteases to <30‐mer peptides is followed by an action of POP. Unexpectedly, POP inhibited the first hydrolysis step, revealing a novel regulation system. POP with Tβ4 significantly induced, while KYP‐2047 effectively prevented, angiogenesis in both models compared with Tβ4 addition itself . POP and endothelial cells were abundantly co‐localized in vivo . CONCLUSIONS AND IMPLICATIONS We have now revealed that POP is a second‐step enzyme in the release of Ac‐SDKP from Tβ4, and it has novel autoregulatory effect in the first step. Our results also advocate a role for Ac‐SDKP in angiogenesis, and suggest that POP has a pro‐angiogenic role via the release of Ac‐SDKP from its precursor Tβ4 and POP inhibitors can block this action.