Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency

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

doi:10.1038/s41598-023-27717-x

Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency

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

Chemistry & Nanoscience

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	1005
Journal	Scientific Reports
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41598-023-27717-x
State	Published - Dec 2023

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Jung, Y. E., Lee, K. W., Cho, J. H., Bae, D. W., Jeong, B. G., Jung, Y. J., Park, S. B., An, Y. J., Kim, K., Lee, G. S., Kang, L. W., Moon, J. H., Lee, J. H., Kim, E. K., Yim, H. S., & Cha, S. S. (2023). Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency. Scientific Reports, 13(1), [1005]. https://doi.org/10.1038/s41598-023-27717-x

@article{1c30650e61eb4cb2a5f530fd2eda1e40,

title = "Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency",

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.",

author = "Jung, {Ye Eun} and Lee, {Kyeong Won} and Cho, {Jae Hyun} and Bae, {Da Woon} and Jeong, {Bo Gyeong} and Jung, {Yeon Ju} and Park, {Soo Bong} and An, {Young Jun} and Kyungchan Kim and Lee, {Ga Seul} and Kang, {Lin Woo} and Moon, {Jeong Hee} and Lee, {Jung Hyun} and Kim, {Eun Kyoung} and Yim, {Hyung Soon} and Cha, {Sun Shin}",

note = "Funding Information: This work was supported by the project entitled “Development of Biomedical materials based on marine proteins” (the Ministry of Oceans and Fisheries, Republic of Korea) and the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIT) (No. NRF2022R1A5A102641311). CD experiments were supported by Korea Basic Science Institute (National Research Facilities and Equipment Center) Grant funded by the Ministry of Education (2020R 1A 6C 101B194). We thank GENUV Inc. for allowing us to use a Biacore 8K system. Funding Information: This work was supported by the project entitled “Development of Biomedical materials based on marine proteins” (the Ministry of Oceans and Fisheries, Republic of Korea) and the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIT) (No. NRF2022R1A5A102641311). CD experiments were supported by Korea Basic Science Institute (National Research Facilities and Equipment Center) Grant funded by the Ministry of Education (2020R 1A 6C 101B194). We thank GENUV Inc. for allowing us to use a Biacore 8K system. Publisher Copyright: {\textcopyright} 2023, The Author(s).",

year = "2023",

month = dec,

doi = "10.1038/s41598-023-27717-x",

language = "English",

volume = "13",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

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T1 - Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency

AU - Jung, Ye Eun

AU - Lee, Kyeong Won

AU - Cho, Jae Hyun

AU - Bae, Da Woon

AU - Jeong, Bo Gyeong

AU - Jung, Yeon Ju

AU - Park, Soo Bong

AU - An, Young Jun

AU - Kim, Kyungchan

AU - Lee, Ga Seul

AU - Kang, Lin Woo

AU - Moon, Jeong Hee

AU - Lee, Jung Hyun

AU - Kim, Eun Kyoung

AU - Yim, Hyung Soon

AU - Cha, Sun Shin

N1 - Funding Information: This work was supported by the project entitled “Development of Biomedical materials based on marine proteins” (the Ministry of Oceans and Fisheries, Republic of Korea) and the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIT) (No. NRF2022R1A5A102641311). CD experiments were supported by Korea Basic Science Institute (National Research Facilities and Equipment Center) Grant funded by the Ministry of Education (2020R 1A 6C 101B194). We thank GENUV Inc. for allowing us to use a Biacore 8K system. Funding Information: This work was supported by the project entitled “Development of Biomedical materials based on marine proteins” (the Ministry of Oceans and Fisheries, Republic of Korea) and the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIT) (No. NRF2022R1A5A102641311). CD experiments were supported by Korea Basic Science Institute (National Research Facilities and Equipment Center) Grant funded by the Ministry of Education (2020R 1A 6C 101B194). We thank GENUV Inc. for allowing us to use a Biacore 8K system. Publisher Copyright: © 2023, The Author(s).

PY - 2023/12

Y1 - 2023/12

N2 - 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.

AB - 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.

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DO - 10.1038/s41598-023-27717-x

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JO - Scientific Reports

JF - Scientific Reports

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M1 - 1005

ER -

Heating-mediated purification of active FGF21 and structure-based design of its variant with enhanced potency

Abstract

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