X-ray crystal structure of plasmin with tranexamic acid–derived active site inhibitors

Ruby H. P. Law, Guojie Wu, Eleanor W. W. Leung, Koushi Hidaka, Adam J. Quek, Tom T. Caradoc-Davies, Devadharshini Jeevarajah, Paul J. Conroy, Nigel M. Kirby, Raymond S. Norton, Yuko Tsuda, James C. Whisstock

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

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 languageEnglish
Pages (from-to)766-771
Number of pages6
JournalBlood Advances
Volume1
Issue number12
DOIs
Publication statusPublished - 9 May 2017

Cite this

@article{8fd45a99e1b54cc5b3d5a55e9f31ee55,
title = "X-ray crystal structure of plasmin with tranexamic acid–derived active site inhibitors",
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.",
author = "Law, {Ruby H. P.} and Guojie Wu and Leung, {Eleanor W. W.} and Koushi Hidaka and Quek, {Adam J.} and Caradoc-Davies, {Tom T.} and Devadharshini Jeevarajah and Conroy, {Paul J.} and Kirby, {Nigel M.} and Norton, {Raymond S.} and Yuko Tsuda and Whisstock, {James C.}",
year = "2017",
month = "5",
day = "9",
doi = "10.1182/bloodadvances.2016004150",
language = "English",
volume = "1",
pages = "766--771",
journal = "Blood Advances",
issn = "2473-9529",
publisher = "American Society of Hematology",
number = "12",

}

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 journalArticleResearchpeer-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 -