Inhibition of human initiator caspase 8 and effector caspase 3 by cross-class inhibitory bovSERPINA3-1 and A3-3

Herrera-Mendez, Carlos H.; Becila, Samira; Blanchet, Xavier; Pelissier, Patrick; Delourme, Didier; Coulis, Gerald; Sentandreu, Miguel A.; Boudjellal, Abdelghani; Bremaud, Laure; Ouali, Ahmed

doi:10.1016/j.febslet.2009.07.055

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Volume 583, Issue 17 , Pages 2743-2748, 3 September 2009

Inhibition of human initiator caspase 8 and effector caspase 3 by cross-class inhibitory bovSERPINA3-1 and A3-3

Edited by Gianni Cesareni

Carlos H. Herrera-Mendez
- UNESS, Universidad de Guanajuato, Privada de Arteaga S/N, 38900 Salvatierra, Guanajuato, Mexico
- These three authors contributed similarly to the present work.
Samira Becila
- INATAA, Université de Constantine, Route de Aïn El Bey, 25000 Constantine, Algeria
- These three authors contributed similarly to the present work.
Xavier Blanchet
- UGMA, UMR 1061, INRA-Université Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cedex, France
- These three authors contributed similarly to the present work.
Patrick Pelissier
- UGMA, UMR 1061, INRA-Université Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cedex, France
Didier Delourme
- UGMA, UMR 1061, INRA-Université Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cedex, France
Gerald Coulis
- UR370, QuaPA, INRA de Clermont Ferrand – Theix, 63122 Saint Genes Champanelle, France
Miguel A. Sentandreu
- Department of Food Science, C.S.I.C., Apt. 73, 46100 Burjassot, Valencia, Spain
Abdelghani Boudjellal
- INATAA, Université de Constantine, Route de Aïn El Bey, 25000 Constantine, Algeria
Laure Bremaud
- UGMA, UMR 1061, INRA-Université Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cedex, France
Ahmed Ouali
- UR370, QuaPA, INRA de Clermont Ferrand – Theix, 63122 Saint Genes Champanelle, France
- Corresponding author. Fax: +33 4 73 62 42 68.

Received 8 June 2009; received in revised form 21 July 2009; accepted 30 July 2009. published online 06 August 2009.

Abstract

Serpins are a superfamily of structurally conserved proteins. Inhibitory serpins use a suicide substrate-like mechanism. Some are able to inhibit cysteine proteases in cross-class inhibition. Here, we demonstrate for the first time the strong inhibition of initiator and effector caspases 3 and 8 by two purified bovine SERPINA3s. SERPINA 3-1 (uniprotkb:Q9TTE1) binds tighly to human CASP3 (uniprotkb:P42574) and CASP8 (uniprotkb:Q14790) with k_ass of 4.2×10⁵ and 1.4×10⁶ M⁻¹s⁻¹, respectively. A wholly similar inhibition of human CASP3 and CASP8 by SERPINA3-3 (uniprotkb:Q3ZEJ6) was also observed with k_ass of 1.5×10⁵ and 2.7×10⁶ M⁻¹s⁻¹, respectively and form SDS-stable complexes with both caspases. By site-directed mutagenesis of bovSERPINA3-3, we identified Asp³⁷¹ as the potential P1 residue for caspases. The ability of other members of this family to inhibit trypsin and caspases was analysed and discussed.