Chemical and thermal unfolding of calreticulin.

Abstract:

:Calreticulin is a soluble endoplasmic reticulum chaperone, which has a relatively low melting point due to its remarkable structure with a relatively high content of flexible structural elements. Using far ultraviolet circular dichroism (CD) spectroscopy and a fluorescent dye binding thermal shift assay, we have investigated the chemical and thermal stability of calreticulin. When the chemical stability of calreticulin was assessed, a midpoint for calreticulin unfolding was calculated to 3.0M urea using CD data at 222 nm. Using the fluorescent dye binding thermal shift assay, calreticulin was found to obtain a molten structure in urea concentrations between 1-1.5 M urea, and to unfold/aggregate at high and low pH values. The results demonstrated that the fluorescent dye binding assay could measure the thermal stability of calreticulin in aqueous buffers with results comparable to melting points obtained by other techniques.

journal_name

Protein Pept Lett

authors

Duus K,Larsen N,Tran TA,Güven E,Skov LK,Jespersgaard C,Gajhede M,Houen G

doi

10.2174/0929866511320050009

subject

Has Abstract

pub_date

2013-05-01 00:00:00

pages

562-8

issue

5

eissn

0929-8665

issn

1875-5305

pii

PPL-EPUB-20120914-1

journal_volume

20

pub_type

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