Abstract
The zymogen protease plasminogen and its active form plasmin perform key roles in blood clot dissolution, tissue remodeling, cell migration, and bacterial pathogenesis. Dysregulation of the plasminogen/plasmin system results in life-threatening hemorrhagic disorders or thrombotic vascular occlusion. Accordingly, inhibitors of this system are clinically important. Currently, tranexamic acid (TXA), a molecule that prevents plasminogen activation through blocking recruitment to target substrates, is the most widely used inhibitor for the plasminogen/plasmin system in therapeutics. However, TXA lacks efficacy on the active form of plasmin. Thus, there is a need to develop specific inhibitors that target the protease active site. Here we report the crystal structures of plasmin in complex with the novel YO (trans-4aminomethylcyclohexanecarbonyl-L-tyrosine-n-octylamide) class of small molecule inhibitors. We found that these inhibitors form key interactions with the S1 and S39 subsites of the catalytic cleft. Here, the TXA moiety of the YO compounds inserts into the primary (S1) specificity pocket, suggesting that TXA itself may function as a weak plasmin inhibitor, a hypothesis supported by subsequent biochemical and biophysical analyses. Mutational studies reveal that F587 of the S9 subsite plays a key role in mediating the inhibitor interaction. Taken together, these data provide a foundation for the future development of small molecule inhibitors to specifically regulate plasmin function in a range of diseases and disorders.
| Original language | English |
|---|---|
| Pages (from-to) | 766-771 |
| Number of pages | 6 |
| Journal | Blood Advances |
| Volume | 1 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 9 May 2017 |
Cite this
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X-ray crystal structure of plasmin with tranexamic acid–derived active site inhibitors. / Law, Ruby H. P.; Wu, Guojie; Leung, Eleanor W. W.; Hidaka, Koushi; Quek, Adam J.; Caradoc-Davies, Tom T.; Jeevarajah, Devadharshini; Conroy, Paul J.; Kirby, Nigel M.; Norton, Raymond S.; Tsuda, Yuko; Whisstock, James C.
In: Blood Advances, Vol. 1, No. 12, 09.05.2017, p. 766-771.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - X-ray crystal structure of plasmin with tranexamic acid–derived active site inhibitors
AU - Law, Ruby H. P.
AU - Wu, Guojie
AU - Leung, Eleanor W. W.
AU - Hidaka, Koushi
AU - Quek, Adam J.
AU - Caradoc-Davies, Tom T.
AU - Jeevarajah, Devadharshini
AU - Conroy, Paul J.
AU - Kirby, Nigel M.
AU - Norton, Raymond S.
AU - Tsuda, Yuko
AU - Whisstock, James C.
PY - 2017/5/9
Y1 - 2017/5/9
N2 - The zymogen protease plasminogen and its active form plasmin perform key roles in blood clot dissolution, tissue remodeling, cell migration, and bacterial pathogenesis. Dysregulation of the plasminogen/plasmin system results in life-threatening hemorrhagic disorders or thrombotic vascular occlusion. Accordingly, inhibitors of this system are clinically important. Currently, tranexamic acid (TXA), a molecule that prevents plasminogen activation through blocking recruitment to target substrates, is the most widely used inhibitor for the plasminogen/plasmin system in therapeutics. However, TXA lacks efficacy on the active form of plasmin. Thus, there is a need to develop specific inhibitors that target the protease active site. Here we report the crystal structures of plasmin in complex with the novel YO (trans-4aminomethylcyclohexanecarbonyl-L-tyrosine-n-octylamide) class of small molecule inhibitors. We found that these inhibitors form key interactions with the S1 and S39 subsites of the catalytic cleft. Here, the TXA moiety of the YO compounds inserts into the primary (S1) specificity pocket, suggesting that TXA itself may function as a weak plasmin inhibitor, a hypothesis supported by subsequent biochemical and biophysical analyses. Mutational studies reveal that F587 of the S9 subsite plays a key role in mediating the inhibitor interaction. Taken together, these data provide a foundation for the future development of small molecule inhibitors to specifically regulate plasmin function in a range of diseases and disorders.
AB - The zymogen protease plasminogen and its active form plasmin perform key roles in blood clot dissolution, tissue remodeling, cell migration, and bacterial pathogenesis. Dysregulation of the plasminogen/plasmin system results in life-threatening hemorrhagic disorders or thrombotic vascular occlusion. Accordingly, inhibitors of this system are clinically important. Currently, tranexamic acid (TXA), a molecule that prevents plasminogen activation through blocking recruitment to target substrates, is the most widely used inhibitor for the plasminogen/plasmin system in therapeutics. However, TXA lacks efficacy on the active form of plasmin. Thus, there is a need to develop specific inhibitors that target the protease active site. Here we report the crystal structures of plasmin in complex with the novel YO (trans-4aminomethylcyclohexanecarbonyl-L-tyrosine-n-octylamide) class of small molecule inhibitors. We found that these inhibitors form key interactions with the S1 and S39 subsites of the catalytic cleft. Here, the TXA moiety of the YO compounds inserts into the primary (S1) specificity pocket, suggesting that TXA itself may function as a weak plasmin inhibitor, a hypothesis supported by subsequent biochemical and biophysical analyses. Mutational studies reveal that F587 of the S9 subsite plays a key role in mediating the inhibitor interaction. Taken together, these data provide a foundation for the future development of small molecule inhibitors to specifically regulate plasmin function in a range of diseases and disorders.
UR - http://www.scopus.com/inward/record.url?scp=85045750082&partnerID=8YFLogxK
U2 - 10.1182/bloodadvances.2016004150
DO - 10.1182/bloodadvances.2016004150
M3 - Article
VL - 1
SP - 766
EP - 771
JO - Blood Advances
JF - Blood Advances
SN - 2473-9529
IS - 12
ER -