Aptamer-peptide conjugates as a new strategy to modulate human α-thrombin binding affinity
Aviñó, Ana (Institut de Química Avançada de Catalunya)
Jorge, Andreia F. (Universidade de Coimbra)
Huertas, César S. (Institut Català de Nanociència i Nanotecnologia)
Cova, Tania F. G. G. (Universidade de Coimbra)
Pais, Alberto (Universidade de Coimbra)
Lechuga, Laura M (Institut Català de Nanociència i Nanotecnologia)
Eritja, Ramon (Institut de Química Avançada de Catalunya)
Fàbrega, Carme (Institut de Química Avançada de Catalunya)

Date:	2019
Abstract:	Aptamers are single-stranded RNA or DNA molecules that specifically recognize their targets and have proven valuable for functionalizing sensitive biosensors. α-thrombin is a trypsin-like serine proteinase which plays a crucial role in haemostasis and thrombosis. An abnormal activity or overexpression of this protein is associated with a variety of diseases. A great deal of attention was devoted to the construction of high-throughput biosensors for accurately detect thrombin for the early diagnosis and treatment of related diseases. Herein, we propose a new approach to modulate the interaction between α-thrombin and the aptamer TBA. To this end, TBA was chemically conjugated to two peptide sequences (TBA-GFIE-Ac and TBA-GEIF-Ac) corresponding to a short fragment of the acidic region of the human factor V, which is known to interact directly with exosite I. Surface Plasmon Resonance (SPR) results showed enhanced analytical performances of thrombin with TBA-GEIF-Ac than with TBA wild-type, reaching a limit of detection as low as 44. 9 pM. Electrophoresis mobility shift assay (EMSA) corroborated the SPR results. Molecular dynamics (MD) simulations support experimental evidences and provided further insight into thrombin/TBA-peptide interaction. Our findings demonstrate that the combination of TBA with key interacting peptides offers good opportunities to produce sensitive devices for thrombin detection and potential candidates to block thrombin activity.
Grants:	Ministerio de Economía y Competitividad CTQ2010-20541 Ministerio de Economía y Competitividad CTQ2014-52588-R Ministerio de Economía y Competitividad CTQ2017-84415-R Agència de Gestió d'Ajuts Universitaris i de Recerca 2009/SGR-208 Ministerio de Economía y Competitividad SEV-2017-0706
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Language:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Subject:	Thrombin ; Biosensor ; Binding affinity ; Inhibition ; Aptamer-peptide conjugate
Published in:	Biochimica et Biophysica Acta - General Subjects, Vol. 1863, Issue 10 (October 2019) , p. 1619-1630, ISSN 1872-8006

DOI: 10.1016/j.bbagen.2019.06.014

Postprint 53 p, 6.1 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > Catalan Institute of Nanoscience and Nanotechnology (ICN2)
Articles > Research articles
Articles > Published articles