Title
Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency
Author
Ye-Eun Jung, Kyeong Won Lee, Jae Hyun Cho, Da-Woon Bae, Bo-Gyeong Jeong, Yeon-Ju Jung, Soo-Bong Park, Young Jun An, Kyungchan Kim, Ga Seul Lee, Lin-Woo Kang, Jeong Hee Moon, Jung-Hyun Lee, Eun-Kyoung Kim, Hyung-Soon Yim & Sun-Shin Cha
Year
2023
Journal
Scientific Reports
Abstract
Fibroblast growth factor 21 (FGF21) has pharmaceutical potential against obesity-related metabolic disorders, including non-alcoholic fatty liver disease. Since thermal stability is a desirable factor for therapeutic proteins, we investigated the thermal behavior of human FGF21. FGF21 remained soluble after heating; thus, we examined its temperature-induced structural changes using circular dichroism (CD). FGF21 showed inter-convertible temperature-specific CD spectra. The CD spectrum at 100 °C returned to that at 20 °C when the heated FGF21 solution was cooled. Through loop swapping, the connecting loop between β10 and β12 in FGF21 was revealed to be associated with the unique thermal behavior of FGF21. According to surface plasmon resonance (SPR) experiments, in vitro cell-based assays, and model high-fat diet (HFD)-induced obesity studies, heated FGF21 maintained biological activities that were comparable to those of non-heated and commercial FGF21s. Based on sequence comparison and structural analysis, five point-mutations were introduced into FGF21. Compared with the wild type, the heated FGF21 variant displayed improved therapeutic potential in terms of body weight loss, the levels of hepatic triglycerides and lipids, and the degree of vacuolization of liver in HFD-fed mice.
Full Article
Instrument
V-730, J-1500
Keywords
FGF21, metabolic, therepeutic proteins, lipid