This article describes the denaturation process of proteins and the application of urea in protein denaturation. Urea and guanidine hydrochloride are strong denaturants that disrupt hydrogen bonding, causing proteins to adopt a highly unfolded and less compact conformation in solution.
This document explains the process of protein denaturation, where proteins adopt a random conformation due to harsh conditions like the addition of urea. It details how chemical denaturants disrupt the non-covalent intramolecular interactions that maintain a protein's tertiary structure, often leading to aggregation.
| Column | Agilent ProSEC 3008, 7.5 x 300 mm |
|---|---|
| Eluent | Containing urea as specified 0.3 M NaCl, pH 8.0 |
| Flow Rate | 1.0 mL/min |
| Injection Volume | 100 pL |
| Temperature | 5°C |
| Detection | UV at 310 nm + Agilent dual angle LSD |
The analysis uses urea at concentrations of 2 M introduced for two hours and 8 M introduced overnight.
Urea instigates protein unfolding, producing higher molecular weight species due to the proteins adopting less compact structures and aggregation through disulfide bond formation.