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@ARTICLE{Mirecka:172157,
author = {Mirecka, Ewa Agnieszka and Gremer, Lothar and Schiefer,
Stephanie and Oesterhelt, Filipp and Stoldt, Matthias and
Willbold, Dieter and Hoyer, Wolfgang},
title = {{E}ngineered aggregation inhibitor fusion for production of
highly amyloidogenic human islet amyloid polypeptide},
journal = {Journal of biotechnology},
volume = {-},
issn = {0168-1656},
address = {Amsterdam [u.a.]},
publisher = {Elsevier Science},
reportid = {FZJ-2014-05667},
pages = {S0168-1656(14)00287-9},
year = {2014},
abstract = {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.},
cin = {ICS-6},
ddc = {540},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {452 - Structural Biology (POF2-452)},
pid = {G:(DE-HGF)POF2-452},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000345684000028},
pubmed = {pmid:24928165},
doi = {10.1016/j.jbiotec.2014.06.006},
url = {https://juser.fz-juelich.de/record/172157},
}