Premium
Effect of detergents on the thermal behavior of elastin‐like polypeptides
Author(s) -
Thapa Arjun,
Han Wei,
Simons Robin H.,
Chilkoti Ashutosh,
Chi Eva Y.,
López Gabriel P.
Publication year - 2013
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.22137
Subject(s) - chemistry , circular dichroism , solubility , chromatography , elastin , solubilization , aqueous solution , membrane , membrane protein , recombinant dna , biochemistry , organic chemistry , medicine , pathology , gene
Elastin‐like polypeptide (ELP) fusions have been designed to allow large‐scale, nonchromatographic purification of many soluble proteins by using the inverse transition cycling (ITC) method; however, the sensitivity of the aqueous lower critical solubility phase transition temperature ( T t ) of ELPs to the addition of cosolutes, including detergents, may be a potential hindrance in purification of proteins with surface hydrophobicity in such a manner. To identify detergents that are known to solubilize such proteins (e.g., membrane proteins) and that have little effect on the T t of the ELP, we screened a number of detergents with respect to their effects on the T t and secondary structures of a model ELP (denoted here as ELP180). We found that mild detergents (e.g., n ‐dodecyl‐β‐ D ‐maltoside, Triton‐X100, and 3‐[(3‐cholamidopropyl) dimethylamino]‐1‐propanesulfonate) do not alter the phase transition behavior or structure (as probed by circular dichroism) of ELP180. This result is in contrast to previous studies that showed a strong effect of other detergents (e.g., sodium dodecylsulfate) on the T t of ELPs. Our results clearly indicate that mild detergents do not preclude ITC‐based separation of ELPs, and thus that ELP fusions may prove to be useful in the purification of detergent‐solubilized recombinant hydrophobic proteins, including membrane proteins, which are otherwise notoriously difficult to extract and purify by conventional separation methods (e.g., chromatography). © 2012 Wiley Periodicals, Inc.