tailieunhanh - Báo cáo khoa học: A facile method for expression and purification of the Alzheimer’s disease-associated amyloid b-peptide

A facile method for expression and purification of the Alzheimer’s disease-associated amyloid b-peptide ´ Dominic M. Walsh1, Eva Thulin2, Aedın M. Minogue1, Niklas Gustavsson3, Eric Pang4, David B. Teplow4 and Sara Linse1,2 1 Laboratory for Neurodegenerative Research, School of Biomolecular and Biomedical Science, Conway Institute, Belfield, University College Dublin, Republic of Ireland 2 Department of Biophysical Chemistry, Chemical Centre, Lund University, Sweden 3 Department of Biochemistry, Chemical Centre, Lund University, Sweden 4 Biopolymer Laboratory, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA Keywords Aß; Alzheimer’s disease; aggregation; amyloid; fibrillogenesis Correspondence D. M. Walsh, Laboratory for Neurodegenerative Research, School of Biomolecular. | A facile method for expression and purification of the Alzheimer s disease-associated amyloid b-peptide Dominic M. Walsh1 Eva Thulin2 Aedín M. Minogue1 Niklas Gustavsson3 Eric Pang4 David B. Teplow4 and Sara Linse1 2 1 Laboratory for Neurodegenerative Research Schoolof Biomolecular and BiomedicalScience Conway Institute Belfield University College Dublin Republic of Ireland 2 Department of BiophysicalChemistry ChemicalCentre Lund University Sweden 3 Department of Biochemistry ChemicalCentre Lund University Sweden 4 Biopolymer Laboratory David Geffen Schoolof Medicine at UCLA Los Angeles CA USA Keywords AB Alzheimer s disease aggregation amyloid fibrillogenesis Correspondence D. M. Walsh Laboratory for Neurodegenerative Research School of Biomolecular and Biomedical Science Conway Institute Belfield University College Dublin Dublin 4 Republic of Ireland Fax 353 1 716 6890 Tel 353 1 716 6751 E-mail S. Linse Department of Biophysical Chemistry Chemical Centre Lund University PO Box 124 SE-22100 Lund Sweden Fax 46 46 2228246 Tel 46 46 224543 E-mail Re-use of this article is permitted in accordance with the Creative Commons Deed Attribution which does not permit commercial exploitation Received 18 November 2008 revised 10 December 2008 accepted 17 December 2008 doi We report the development of a high-level bacterial expression system for the Alzheimer s disease-associated amyloid b-peptide Ab together with a scaleable and inexpensive purification procedure. Ab 1-40 and AP 1 42 coding sequences together with added ATG codons were cloned directly into a Pet vector to facilitate production of Met-Ab 1-40 and Met-AP 1-42 referred to as Ab M1-40 and Ab M1-42 respectively. The expression sequences were designed using codons preferred by Escherichia coli and the two peptides were expressed in this host in inclusion bodies. Peptides were purified from inclusion bodies using a combination of .