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Updated: Aug 16, 2026

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells
Published on: June 2, 2022
Sequential conformation changes of chinese hamster ovary dihydrofolate reductase at its active sites
Wen-he Zhang1, San-bo Qin, Hong-jie Zhang
1Institute of Biophysics, Academy Sinica, 15 Datun Road, Beijing 100101, China.
Abstract:
The local fluorescence probes, 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) and NADPH were employed to detect urea-induced conformation changes at each active site of dihydrofolate reductase (DHFR), respectively. The results indicate that local conformation change at DHF/TNS could be superimposed by the conformation change calculated from the enzyme activity change with a three-state model; while at NADPH site it is lagged in the first transition. This difference is further supported by the different relative changes of Michaelis constants at 0, 1 and 1.8 M urea for each substrate. Our results suggest that local conformation at DHF site is more flexible than that at NADPH site, and the urea-induced unfolding could be ascribed to a four-state transition.
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