@article{10902/37403,
year = {2025},
url = {https://hdl.handle.net/10902/37403},
abstract = {Haematophagous organisms are a rich source of salivary anticoagulant polypeptides that exert their activity by blocking the catalytic site and one of two positively charged exosites on the host protease thrombin. Here, we describe a molecular engineering approach to hybridise post-translationally sulfated polypeptides from different blood-feeding organisms to enhance anticoagulant activity. This led to the discovery of a triply sulfated hybrid anticoagulant, XChimera, possessing fragments from flea, leech, and fly salivary polypeptides that exhibits femtomolar inhibitory activity against thrombin. The crystallographic structure of a complex of XChimera with thrombin shows that it displays a trivalent binding mode in which it simultaneously blocks three functional sites of the protease, the active site and exosites I and II. This trivalent chimera exhibited ultrapotent anticoagulant activity in a suite of in vitro clotting assays and was also shown to possess potent in vivo antithrombotic activity in a murine model of thrombosis.},
organization = {This work was funded through National Health and Medical Research Council of Australia Investigator Grants (APP1174941; to R. J. P.; APP1176016; to S. P. J.) and was also supported in part by national funds through FCT – Fundaç˜ao para a Ciˆencia e a Tecnologia, I. P. (Portugal) under project UIDB/04293/2020 and in the scope of research grants PTDC/BIA-BQM/2494/2020 (https://doi.org/10.54499/PTDC/BIA-BQM/2494/2020) and 2022.03363.PTDC (https://doi.org/10.54499/2022.03363.PTDC); and a New South Wales (NSW, Australia) Ministry of Health (MOH) Cardiovascular Senior Researcher Grants awarded to S. M. S. J. R.-R. acknowledges the support of grant RYC2021- 033063-I funded by MCIN/AEI/10.13039/501100011033 and the European Union «NextGenerationEU»/PRTR. X-ray data collection was performed at BL13-XALOC beamline of ALBA Synchrotron with the collaboration of ALBA staff. The support of the X-Ray Crystallography, Biochemical and Biophysical Technologies and BioSciences Screening platforms of i3S (Porto, Portugal) is also acknowledged.},
publisher = {Royal Society of Chemistry},
publisher = {Chemical Science, 2025, 4734J, 1-28},
title = {Engineering ultrapotent trivalent anticoagulants through hybridisation of salivary peptides from multiple haematophagous organisms},
author = {Maxwell, Joshua W. C. and Ripoll Rozada, Jorge and Mackay, Angus S. and Alwis, Imala and Ford, Daniel J. and Trought, Cameron B. J. and Santos, Joana A. and Smythe, Rhyll e. and Liu, Joanna S. T. and Zuccolotto, Zack and Schoenwaelder, Simone M. and Jackson, Shaun P. and Barbosa Pereira, Pedro José and Payne, Richard J.},
}