Surfactants modify the torsion properties of proteins: a single molecule study

• Published in: New biotechnology Vol. 32; no. 5; pp. 441 - 449
• Main Authors: Gutiérrez-Mejía, F.A., van IJzendoorn, L.J., Prins, M.W.J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25-09-2015
• Subjects: Circular Dichroism; Micelles; Protein Conformation; Proteins - chemistry; Sodium Dodecyl Sulfate - chemistry; Spectrophotometry, Ultraviolet; Surface-Active Agents - chemistry
• ISSN: 1871-6784; 1876-4347
• DOI: 10.1016/j.nbt.2015.02.005

•Magnetic tweezers show increased twisting of proteins to surfactant exposure.•Tween 20 decreases the torsional rigidity of protein-G–IgG more than SDS.•Far-UV and near-UV Circular Dichroism do not show strong denaturation of protein-G and IgG.•Magnetic tweezers are sensitive to local changes of the binding between proteins.
Surfactants are widely used in diagnostic assays to prevent protein aggregation and non-specific adsorption at surfaces. Here, a single molecule magnetic torque tweezers study is reported, aiming to quantify surfactant-induced changes in the torsional flexibility of a protein model system: protein-G–immunoglobulin G (IgG) attached to a glass surface. The influences of Sodium Dodecyl Sulphate (SDS) and Polysorbate 20 (Tween 20) on the protein pair have been investigated. The proteins were exposed to the surfactants at concentrations relative to the Critical Micelle Concentration (CMC), namely 0.1× CMC, 1× CMC and 10× CMC. Both surfactants increase the torsional flexibility of the protein-G–IgG complex. Tween 20 is most effective at increasing the torsional flexibility of the complex at the surface while SDS is more effective at dissociating the protein bonds. Tweezer data on the IgG–IgG protein pair show no influence of Tween 20 on the torsional flexibility. Furthermore, temperature dependent near-UV and far-UV Circular Dichroism (CD) data at 10× CMC show that Tween 20 does not significantly alter the secondary and tertiary structure of both protein-G and IgG while SDS does. These results provide evidence that both the mechanical properties of the protein structure and the interaction between proteins can alter the torsional rigidity measured with magnetic torque tweezers. This study shows for the first time the ability to use magnetic torque tweezers as a probe for surfactant-induced changes in proteins at a single molecule level.