Data on solubilization, identification, and thermal stability of human Presenilin-2

Abstract

The data presented here are related to the research article entitled “Expression, purification, and preliminary characterization of human presenilin-2" [1].Human Presenilin-2 is the catalytic subunit of γ-secretase and a possible calcium leakage channel (Kimberly et al., 2000; Tu et al., 2006) [2], [3]. HisPS2 which was obtained by overexpression in E. coli strain C43 (DE3) was extracted by detergent solubilisation. The sample isolation efficiency by detergents and the protein identification by mass spectrometry and western blot are described.This data article describes the near and far UV circular dichroism measurements and the data deconvolution in terms of secondary structure at 4 and 98 °C. Also, a refolding spectrum is presented.The raw CD spectra used for deconvolution of the helix and stand segments and average length are deposited into Protein Circular Dichroism Data Bank with PCDDBid: CD0005962000 (4 °C far UV), CD0005963000 (98 °C far UV), CD0005964000 (back to 4 °C far UV) and CD0005965000 (4 °C near UV CD).

Expression, purification, and preliminary characterization of human presenilin-2

“…All the peptides detected belong to extra-membrane segments (Fig. 2 in Ref [54]), which is similar to the MS result reported for PS1 [62]. The low sequence coverage in MS is probably a result of the inaccessibility of the cleavage sites due to bound detergent.…”

“…The detergents used in the study are indicated in Table 1 in Ref. [54]. After solubilization, the supernatant and pellet were separated by centrifugation at 100,000g for 1 h and analyzed by western blot.…”

“…After overexpression, we investigated the ability of 13 detergents with different soluble groups, charge, and chain length to solubilize hisPS2 (Table 1 in Ref [54]). After detergent extraction (Fig.…”

“…The two dominant bands were found to contain PS2 by MS based on five identified peptides ( Fig. 1 in Ref [54]) with 14% sequence coverage (Fig. 3b).…”

“…Each lane was loaded with an amount corresponding to 3.4 mg of cell pellet. S: suspension before 100,000 × g; SN: supernatant after 100,000 × g; P: pellet after 100,000 × g. The full name of detergents are listed in Table 1 in Ref [54]. N-terminal his-tag was investigated by monoclonal anti-polyhistidine-HRP antibody (Sigma-Aldrich, Hamburg, Germany).…”