Related Experiment Video
Updated: Jul 5, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction
Published on: January 8, 2019
Determining the identity and structure of recombinant proteins
N D Denslow1, K Rose, P G Righetti
1University of Florida, Gainesville, Floria, USA.
Abstract:
In this unit peptide mapping protocols with separation of the constituent peptides by high-performance liquid chromatography (HPLC) analysis and by high-resolution SDS-PAGE are presented. Peptide mapping is ideally suited for comparative purposes--for example, combined analysis of the recombinant protein and its natural counterpart (or some other well-characterized standard). This unit also outlines the general strategy used to determine the linkage pattern of a monomeric recombinant protein containing two intramolecular disulfide bonds. The approach is an extension of peptide mapping, where the aim is to isolate and characterize peptides containing only a single disulfide bond. A two-dimensional electrophoretic method is also described in which the protein isoelectric point is displayed as a function of pH to yield an electrophoretic titration curve. This method is especially useful for checking for deamidation (e.g., of Asn to Asp) in which additional negative charge is introduced into the modified protein.
Related Concept Videos
Recombinant DNA
Tagging and Fusion Proteins
Protein Folding
Protein Folding
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein and Protein Structure
A protein's shape is critical to its function. For example, an enzyme can...
Production of Pharmaceuticals

