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Title: Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state

Abstract

Highlights: Black-Right-Pointing-Pointer CotA-laccase unfolds with an intermediate state. Black-Right-Pointing-Pointer Copper stabilizes the native and the intermediate state. Black-Right-Pointing-Pointer Copper binding to the unfolded state prevents refolding through protein aggregation. Black-Right-Pointing-Pointer Copper incorporation in CotA-laccase occurs as a later step during folding. -- Abstract: Copper is a redox-active metal and the main player in electron transfer reactions occurring in multicopper oxidases. The role of copper in the unfolding pathway and refolding of the multicopper oxidase CotA laccase in vitro was solved using double-jump stopped-flow experiments. Unfolding of apo- and holo-CotA was described as a three-state process with accumulation of an intermediate in between the native and unfolded state. Copper stabilizes the native holo-CotA but also the intermediate state showing that copper is still bound to this state. Also, copper binds to unfolded holo-CotA in a non-native coordination promoting CotA aggregation and preventing refolding to the native structure. These results gather information on unfolding/folding pathways of multicopper oxidases and show that copper incorporation in vivo should be a tight controlled process as copper binding to the unfolded state under native conditions promotes protein aggregation.

Authors:
 [1];  [2];  [1];  [2]
  1. Instituto de Tecnologia Quimica e Biologica, Universidade Nova de Lisboa, Av. da Republica, 2780-157 Oeiras (Portugal)
  2. Centre for Molecular and Structural Biomedicine, Institute for Biotechnology and Bioengineering, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro (Portugal)
Publication Date:
OSTI Identifier:
22207881
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 422; Journal Issue: 3; Other Information: Copyright (c) 2012 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AGGLOMERATION; COPPER; ELECTRON TRANSFER; IN VITRO; IN VIVO; INTERMEDIATE STATE; OXIDASES

Citation Formats

Fernandes, Andre T., Lopes, Carlos, Martins, Ligia O., and Melo, Eduardo Pinho, E-mail: emelo@ualg.pt. Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state. United States: N. p., 2012. Web. doi:10.1016/J.BBRC.2012.05.011.
Fernandes, Andre T., Lopes, Carlos, Martins, Ligia O., & Melo, Eduardo Pinho, E-mail: emelo@ualg.pt. Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state. United States. doi:10.1016/J.BBRC.2012.05.011.
Fernandes, Andre T., Lopes, Carlos, Martins, Ligia O., and Melo, Eduardo Pinho, E-mail: emelo@ualg.pt. Fri . "Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state". United States. doi:10.1016/J.BBRC.2012.05.011.
@article{osti_22207881,
title = {Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state},
author = {Fernandes, Andre T. and Lopes, Carlos and Martins, Ligia O. and Melo, Eduardo Pinho, E-mail: emelo@ualg.pt},
abstractNote = {Highlights: Black-Right-Pointing-Pointer CotA-laccase unfolds with an intermediate state. Black-Right-Pointing-Pointer Copper stabilizes the native and the intermediate state. Black-Right-Pointing-Pointer Copper binding to the unfolded state prevents refolding through protein aggregation. Black-Right-Pointing-Pointer Copper incorporation in CotA-laccase occurs as a later step during folding. -- Abstract: Copper is a redox-active metal and the main player in electron transfer reactions occurring in multicopper oxidases. The role of copper in the unfolding pathway and refolding of the multicopper oxidase CotA laccase in vitro was solved using double-jump stopped-flow experiments. Unfolding of apo- and holo-CotA was described as a three-state process with accumulation of an intermediate in between the native and unfolded state. Copper stabilizes the native holo-CotA but also the intermediate state showing that copper is still bound to this state. Also, copper binds to unfolded holo-CotA in a non-native coordination promoting CotA aggregation and preventing refolding to the native structure. These results gather information on unfolding/folding pathways of multicopper oxidases and show that copper incorporation in vivo should be a tight controlled process as copper binding to the unfolded state under native conditions promotes protein aggregation.},
doi = {10.1016/J.BBRC.2012.05.011},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 3,
volume = 422,
place = {United States},
year = {2012},
month = {6}
}