TY  - JOUR
AU  - Yang, Ge
AU  - Yu, Kun
AU  - Kubicek, Jan
AU  - Labahn, Jörg
TI  - Expression, purification, and preliminary characterization of human presenilin-2
JO  - Process biochemistry
VL  - 64
SN  - 1359-5113
CY  - Amsterdam [u.a.]
PB  - Elsevier Science
M1  - FZJ-2017-06601
SP  - 63-73
PY  - 2018
AB  - Presenilins (PS1 and PS2) exhibit similar γ-secretase-dependent and −independent functions with subtle variations. In this study, we established a cost-effective process to overexpress and purify full-length human PS2 in sufficient quantities and quality for structural studies. Upon optimization, milligram quantities of homogeneous trimeric hisPS2 were purified, which enabled the preliminary characterization of human hisPS2 zymogen. Far-UV and near-UV CD as well as fluorescence spectroscopy revealed that purified hisPS2 contained the expected secondary structure and was folded into a defined tertiary structure. Thermal stability analysis revealed a Tm value of ∼55 °C for secondary structure while cholesterol significantly increased the stability. The low melting temperature of ∼34 °C for the tertiary structure was able to explain the purity and aggregation problems observed during purification. Additionally, the occurrence of calcium ions induced structural changes to different extents for PS2WT and PS2-D263A/D366A was observed, which is consistent with previous studies.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000423003200008
DO  - DOI:10.1016/j.procbio.2017.09.012
UR  - https://juser.fz-juelich.de/record/837812
ER  -