Adipocyte-derived leucine aminopeptidase (A-LAP) is a novel member of the M1 family of zinc metallopeptidases, which has been reported to play a crucial role in angiogenesis. In the present study, we conducted a target-based screening of natural products and synthetic chemical libraries using the purified enzyme to search novel inhibitors of A-LAP. Amongst several hits isolated, a natural product purpurin was identified as one of the most potent inhibitors of A-LAP from the screening. In vitro enzymatic analyses demonstrated that purpurin inhibited A-LAP activity in a non-competitive manner with a Ki value of 20 M. In addition, purpurin showed a strong selectivity toward A-LAP versus another member of M1 family of zinc metallopeptidase, aminopeptidase N (APN). In angiogenesis assays, purpurin inhibited the vascular endothelial growth factor (VEGF)-induced invasion and tube formation of human umbilical vein endothelial cells (HUVEC). Moreover, purpurin inhibited in vivo angiogenesis in zebrafish embryo without toxicity. These data demonstrate that purpurin is a novel specific inhibitor of A-LAP and could be developed as a new anti-angiogenic agent.
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|Publication status||Published - 2014 Jul 18|
Bibliographical noteFunding Information:
This work was supported by National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIP; 2009-0092964 , 2009-0083522 , 2010-0017984 , 2012M3A9D1054520 ), the Center for Food and Drug Materials of Agriculture Science & Technology Development ( PJ0079772012 ), Rural Development Administration, National R&D Program, Ministry of Health &Welfare ( 0620360-1 ), and the Brain Korea 21 Plus Project, Republic of Korea .
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology