Protocol for reconstituting peptides/peptidomimetics from DMSO to aqueous buffers for circular dichroism analyses.

William H Deni, Tong Gao, Jinhua Wu
Author Information
  1. William H Deni: Cancer Signaling and Microenvironment Program, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
  2. Tong Gao: Cancer Signaling and Microenvironment Program, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
  3. Jinhua Wu: Cancer Signaling and Microenvironment Program, Fox Chase Cancer Center, Philadelphia, PA 19111, USA. Electronic address: jinhua.wu@fccc.edu.

Abstract

Circular dichroism (CD) spectrometry is a rapid technique for detecting protein secondary structure, particularly helicity. DMSO is used to ensure optimal solubility of peptides/peptidomimetics; however, its background absorbance hinders effective CD analysis. Here, we present a protocol for reconstituting peptides/peptidomimetics from DMSO to aqueous buffers for CD analyses. We describe steps for identifying chemicals that induce DMSO evaporation, extracting peptides/peptidomimetics from DMSO, and CD spectrometer setup and analysis. We then detail procedures for secondary structure analyses of reconstituted peptides/peptidomimetics. For complete details on the use and execution of this protocol, please refer to Gao et al. (2023)..

Keywords

Grants

  1. R35 GM119560/NIGMS NIH HHS

MeSH Term

Circular Dichroism
Dimethyl Sulfoxide
Peptidomimetics
Peptides
Proteins
Water

Chemicals

Dimethyl Sulfoxide
Peptidomimetics
Peptides
Proteins
Water

Word Cloud

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