%0 Journal Article
%A Mirecka, Ewa Agnieszka
%A Gremer, Lothar
%A Schiefer, Stephanie
%A Oesterhelt, Filipp
%A Stoldt, Matthias
%A Willbold, Dieter
%A Hoyer, Wolfgang
%T Engineered aggregation inhibitor fusion for production of highly amyloidogenic human islet amyloid polypeptide
%J Journal of biotechnology
%V -
%@ 0168-1656
%C Amsterdam [u.a.]
%I Elsevier Science
%M FZJ-2014-05667
%P S0168-1656(14)00287-9
%D 2014
%X Human islet amyloid polypeptide (IAPP) is the major component of pancreatic amyloid deposits in type 2 diabetes. The structural conversion of IAPP from a monomeric state into amyloid assemblies is the subject of intense research. Recombinant production of IAPP is, however, difficult due to its extreme aggregation propensity. Here we describe a novel strategy for expression of IAPP in Escherichia coli, based on an engineered protein tag, which sequesters IAPP monomers and prevents IAPP aggregation. The IAPP-binding protein HI18 was selected by phage display from a β-wrapin library. Fusion of HI18 to IAPP enabled the soluble expression of the construct. IAPP was cleaved from the fusion construct and purified to homogeneity with a yield of 3mg of isotopically labeled peptide per liter of culture. In the monomeric state, IAPP was largely disordered as evidenced by far-UV CD and liquid-state NMR spectroscopy but competent to form amyloid fibrils according to atomic force microscopy. These results demonstrate the ability of the engineered β-wrapin HI18 for shielding the hydrophobic sequence of IAPP during expression and purification. Fusion of aggregation-inhibiting β-wrapins is a suitable approach for the recombinant production of aggregation-prone proteins.
%F PUB:(DE-HGF)16
%9 Journal Article
%U <Go to ISI:>//WOS:000345684000028
%$ pmid:24928165
%R 10.1016/j.jbiotec.2014.06.006
%U https://juser.fz-juelich.de/record/172157