1. Substrate specificity of a metalloprotease of the pappalysin family revealed by an inhibitor and a product complex
- Author
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Cynthia Tallant, Maria Solà, Aniebrys Marrero, Ulrich Baumann, F. Xavier Gomis-Rüth, and Raquel García-Castellanos
- Subjects
Models, Molecular ,Metzincin ,Protein Folding ,Stereochemistry ,Phenylalanine ,medicine.medical_treatment ,Molecular Sequence Data ,Biophysics ,Thiophenes ,Biology ,Matrix metalloproteinase ,Crystallography, X-Ray ,Biochemistry ,Substrate Specificity ,Ulilysin ,Pappalysin ,Metalloprotease ,chemistry.chemical_compound ,Astacin ,Catalytic Domain ,Hydrolase ,medicine ,Batimastat ,Metalloprotease inhibitor ,Amino Acid Sequence ,IGF ,Molecular Biology ,Cancer ,Metalloproteinase ,Hydroxamic acid ,Protease ,Sequence Homology, Amino Acid ,X-ray crystal structure ,Serralysin ,Enzyme-inhibitor complex ,chemistry ,Metalloproteases ,Oligopeptides ,Protein Binding ,IGFBP protease - Abstract
Human pappalysin-1 is a multi-domain metalloprotease engaged in the homeostasis of insulin-like growth factors and the founding member of the pappalysin family within the metzincin clan of metalloproteases. We have recently identified an archaeal relative, ulilysin, encompassing only the protease domain. It is a 262-residue active protease with a novel 3D structure with two subdomains separated by an active-site cleft. Despite negligible overall sequence similarity, noticeable similarity is found with other metzincin prototypes, adamalysins/ADAMs and matrix metalloproteinases. Ulilysin has been crystallised in a product complex with an arginine-valine dipeptide occupying the active-site S1′ and S2′ positions and in a complex with the broad-spectrum hydroxamic acid-based metalloprotease inhibitor, batimastat. This molecule inhibits mature ulilysin with an IC50 value of 61 μM under the conditions assayed. The binding of batimastat to ulilysin evokes binding to vertebrate matrix metalloproteases but is much weaker. These data give insight into substrate specificity and mechanism of action and inhibition of the novel pappalysin family. © 2006 Elsevier Inc. All rights reserved., This study was supported by the following grants and fellowships: SAB2002-0102 from the former Spanish Ministry for Education, Culture and Sports; BIO2003-00132, GEN2003-20642 and BIO2004-20369-E from the former Spanish Ministry for Science and Technology; BIO2006-02668 and BFU2006-09593 and CONSOLIDER-INGENIO 2010 Project ‘La Factorı´a de Cristalización’ (CSD2006-00015) from the Spanish Ministry for Education and Science; ON03-7-0 from the “Fundació La Caixa”; EU FP6 Integrated Project LSHC-CT-2003-503297 “CANCERDEGRADOME”; “AVON-Project” 2005X0648 from the Scientific Foundation of the Spanish Association Against Cancer; Grant 2005SGR00280 from the Generalitat of Catalunya; and grants 31-67253.01 and 3100AO-108262/1 from the Swiss National Science Foundation. M.S. is a beneficiary of the “Ramon y Cajal” Program from the Spanish Ministry for Science and Education
- Published
- 2007
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