Studies on the peptidase activity of transthyretin (TTR)

Studies on the peptidase activity of transthyretin (TTR)

Author Gouvea, Iuri Estrada Autor UNIFESP Google Scholar
Kondo, Marcia Yuri Autor UNIFESP Google Scholar
Assis, Diego M. Autor UNIFESP Google Scholar
Alves, Fabiana Madureira Autor UNIFESP Google Scholar
Liz, Marcia Almeida Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Inst Biol Mol & Celular
Abstract Transthyretin (TTR) is a plasma protein transporter of thyroxine (T-4) and retinol and also has peptidase activity. in order to characterize TTR peptidase activity we used fluorescence resonance energy transfer (FRET) peptides derived from Abz-KLRSSK-Q-EDDnp and from two portion-mixing libraries as substrates. Most of the susceptible FRET peptides were cleaved at more than one peptide bond, without particular substrate specificity. the more relevant observation was that the peptides containing E or D were cleaved at only one peptide bond and TTR was competitively inhibited by glutathione analog peptide gamma-E-A-G-OH that contains two free carboxyl groups. the dependence on ionic interactions of TTR hydrolytic activity was confirmed by the large inhibitory effects of salt and ionic surfactants. TTR was not inhibited by any usual peptidase inhibitors, except by ortho-phenanthroline and EDTA. the mechanism of TTR catalysis was explored by the pH-profile of TTR hydrolytic activity in different temperatures and by proton inventory. the obtained pK and heat of ionization values suggest that a carboxylate and an ammonium group, possibly from a lysine side chain are involved. These results support the recently proposed inducible metalloprotease mechanism for TTR based on its 3D structure in presence of Zn2+ and a series of point mutations [Liz et al., Biochem. J 443 (2012) 769-778]. (c) 2012 Elsevier Masson SAS. All rights reserved.
Keywords Transthyretin
Protease
Fluorescent peptides
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundacao para a Ciencia e Tecnologia under the program FEDER in Portugal
Fundacao para a Ciencia e Tecnologia under the program COMPETE in Portugal
Grant number Fundacao para a Ciencia e Tecnologia under the program FEDER in Portugal: PTDC/SAU-GMG/111761/2009
Fundacao para a Ciencia e Tecnologia under the program FEDER in Portugal: PTDC/SAU-ORG/118863/2010
Fundacao para a Ciencia e Tecnologia under the program COMPETE in Portugal: PTDC/SAU-GMG/111761/2009
Fundacao para a Ciencia e Tecnologia under the program COMPETE in Portugal: PTDC/SAU-ORG/118863/2010
Date 2013-02-01
Published in Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 95, n. 2, p. 215-223, 2013.
ISSN 0300-9084 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 215-223
Origin http://dx.doi.org/10.1016/j.biochi.2012.09.014
Access rights Closed access
Type Article
Web of Science ID WOS:000315614200010
URI http://repositorio.unifesp.br/handle/11600/35926

Show full item record




File

File Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Search


Browse

Statistics

My Account