Protein Folding Assisted by Covalent Hydrogel Based on Amphiphilic Supramolecular

doi:10.7503/cjcu20230250

Abstract

Abstract:

A novel monomeric species， 2-｛6-［3-（6-methyl-4-oxo-1，4-dihydropyrimidin-2-yl）ureido］hexylcarbamoyloxy｝ butyl acrylate（UPy-C₆-HBA）， was synthesized while 4-hydroxybutyl acrylate（HBA） was introduced to the intermediate compound 2-（6-isocyanatohexyl ureido）-6-methyl-4［1H］ pyrimidinone（UPy-C₆-NCO） which was obtained through the reaction of 2-aminomethyl-4-hydroxy-6-methylpyrimidine（MIS） with 1，6-hexane diisocyanate（HDI）. UPy-C₆-HBA together with acrylamide（AM） and N，N'-methylenediacrylamide（Bis） was polymerized by UV-induced free radical polymerization method， yielding physically crosslinked hydrogels that possessed a homogenous porous architecture and pronounced swelling capacity. Lysozyme was incorporated within the hydrogel matrix containing UPy-C₆-HBA monomer which in vitro refolding was facilitated via hydrophobic interactions. The effects of UPy-C₆-HBA concentration， hydrogel dosage， lysozyme concentration and refolding temperature on the refolding rate of lysozyme were meticulously investigated. Remarkably， the supramolecular hydrogel comprising UPy-C₆-HBA at a mass fraction of 15% exhibited a remarkable 41% enhancement in the refolding rate of lysozyme at 25 ℃ compared with the control. These findings unequivocally demonstrate the addition of UPy-C6-HBA monomer to the hydrogel matrix significantly improved the refolding rate of lysozyme， which is of great significance for the refolding of high concentration denatured lysozyme.

Key words: 2-｛6-［3-（6-Methyl-4-oxo-1, 4-dihydropyrimidin-2-yl）ureido］hexylcarbamoyloxy｝butyl acrylate, Supramolecular dual-crosslinked hydrogels, Protein folding, Lysozyme

CLC Number:

O633

TrendMD:

WANG Jiaxin, LIU Jia, QIN Jinmei, NYV Mondele Mbola, MENG Zihui, XUE Min. Protein Folding Assisted by Covalent Hydrogel Based on Amphiphilic Supramolecular[J]. Chem. J. Chinese Universities, 2023, 44(11): 20230250.

Figures/Tables 9

References 21

1	Bolen D. W.， Rose G. D.， Annual Review of Biochemistry， 2008， 77（1）， 339—362
2	Su L. L.， Shao X. G.， Cai W. S.， Chem. J. Chinese Universities， 2023， 44（4）， 20220745
	粟李醴，邵学广，蔡文生. 高等学校化学学报， 2023， 44（4）， 20220745
3	Curtis R.， Austerberry J.， Holloway L.， Compr. Biotechnol， 2019， 2， 860—878
4	Yamaguchi H.， Miyazaki M.， Biomolecules， 2014， 4（1）， 235—251
5	Wang Y.， Oosterwijk N. V.， Ali A. M.， Adawy A.， Anindya A. L.， Dömling A. S. S.， Groves M. R.， Sci. Rep.， 2017， 7（1）， 9345—9355
6	Wang X.， Lu D.， Austin R.， Agarwal A.， Mueller L. J.， Liu Z.， Wu J.， Feng P.， Langmuir， 2007， 23（10）， 5735—5739
7	Lu J.， Liong M.， Li Z.， Zink J. I.， Tamanoi F.， Small， 2010， 6（16）， 1794—1805
8	Singh N.， Nisha V.， Am. J. Adv. Drug Deliv.， 2013， 1， 271—276
9	Nakamoto M.， Nonaka T.， Shea K. J.， Miura Y.， Hoshino Y.， J. Am. Chem. Soc.， 2016， 138（13）， 4282—4285
10	Tandon S.， Horowitz P. M.， J. Biol. Chem.， 1987， 262（10）， 4486—4491
11	Kameta N.， Masuda M.， Shimizu T.， ACS Nano， 2012， 6（6）， 5249—5258
12	Skopinska W.， Flor J. S.， Kozlowska J.， Int. J. Mol. Sci.， 2021， 22（14）， 7402
13	Shi Z.， Wang Q.， Li G. F.， Shou Y. F.， Zong H. J.， Yan S. F.， Zhang K. X.， Yin J. B.， J. Polym. Sci.， 2021， 39（3）， 327—336
14	Lim J. Y. C.， Lin Q.， Xue K.， Loh X. J.， Mater. Today Adv.， 2019， 3， 100021
15	Mol E. A.， Lei Z.， Roefs M. T.， Bakker M. H.， Goumans M. J.， Doevendans P. A.， Dankers P. Y. W.， Vader P.， Sluijter J. P. G.， Adv. Healthc. Mater.， 2019， 8（20）， e1900847
16	Balavigneswaran C. K.， Muthuvijayan V.， ACS Appl. Bio. Mater.， 2021， 4（6）， 5362—5377
17	Mes T.， Serrero A.， Bauer H. S.， Cox M. A. J.， Bosman A. W.， Dankers P. Y. W.， Meijer E. W.， Mater. Today， 2022， 52， 175—187
18	Webber M. J.， Dankers P. Y. W.， Macromol. Biosci.， 2019， 19（1）， e1800452
19	Gao T. T.， Li Z. Y.， Gao G.， Liu F. Q.， Chem. J. Chinese Universities， 2016， 37（9）， 1744—1749
	高婷婷，李志英，高歌，刘凤岐. 高等学校化学学报， 2016， 37（9）， 1744—1749
20	Ma S. H.， An Y. L.， Wang J. Z.， Song Y. Q.， Liu Y.， Shi L. Q.， ACS Nano， 2017， 11（10）， 10549—10557
21	Takeda S.， Takahashi H.， Sawada S.， Sasaki Y.， Akiyoshi K.， RSC Adv.， 2013， 3， 25716

[1]	SU Lili, SHAO Xueguang, CAI Wensheng. Mechanism of Tau R2/Wt Morphology Affected by Osmolytes Using Molecular Dynamics Simulations [J]. Chem. J. Chinese Universities, 2023, 44(4): 20220745.
[2]	WU Yao, LIANG Wenjing, LI Cheng, SHANG Xu, CONG Lina. Effects of Temperature on the Antibacterial Activity and Structural Change of the C-terminal Polypeptide of the Sea Cucumber Lysozyme^† [J]. Chem. J. Chinese Universities, 2014, 35(5): 1044.
[3]	FENG Xuan, ZHANG Tan, BIAN Liu-Jiao. Distribution and Transition of Stable Conformations of Denatured Hen Egg White Lysozymes During Their Refolding in Guanidine Hydrochloride Solutions [J]. Chem. J. Chinese Universities, 2013, 34(2): 336.
[4]	KE Cong-Yu, MENG Zu-Chao. Denatured Ribonuclease Refolding by Glutathione Bonding Column [J]. Chem. J. Chinese Universities, 2012, 33(05): 925.
[5]	WU Xian-Li, HAN Guo-Sheng, WANG Zhen, CAO Shu-Xia, LIAO Xin-Cheng. Interaction Between N-Phosphoryl Dopamines and Lysozyme by ESI-MS [J]. Chem. J. Chinese Universities, 2012, 33(03): 516.
[6]	ZHANG Tan, BIAN Liu-Jiao*. Distribution and Transition of Stable Conformations of Hen Egg White Lysozymes in Their Unfolding Induced by Urea and Guanidine Hydrochloride Solutions [J]. Chem. J. Chinese Universities, 2011, 32(7): 1497.
[7]	HUANG Shan, XIAO Qi, HE Zhi-Ke*, LIU Yi. [J]. Chem. J. Chinese Universities, 2009, 30(10): 1951.
[8]	YU Shao-Yong, GAO Gao, YAO Ping*, JIANG Ming. Loading and Release of Lysozyme by Casein-g-dextran Graft Copolymer [J]. Chem. J. Chinese Universities, 2008, 29(5): 1027.
[9]	DU Zhuo, ZHANG Na, WANG Jian-Hua*. On-line Isolation of Lysozyme from Egg-white Using Multi-walled Carbon Nanotubes [J]. Chem. J. Chinese Universities, 2008, 29(5): 902.
[10]	LIU Qiu-Ye¹, GAI Qing-Qing¹, HE Xi-Wen¹, LI Wen-You¹, CHEN Lang-Xing¹, ZHANG Yu-Kui^1,2. Matrix Molecularly Imprinted Polymer for Enriching Lysozyme in Protein Samples [J]. Chem. J. Chinese Universities, 2008, 29(3): 505.
[11]	YANG Ran¹, CHEN Xiao-Lan², LI Ping¹, QU Ling-Bo². Interaction Between Lysozyme and Three Flavones by Fluorescence Spectroscopy [J]. Chem. J. Chinese Universities, 2006, 27(9): 1673.
[12]	WANG Chun-Yan^1,2, ZHU Guang-Shan¹, GAO Bo³, WANG Run-Wei¹, LI Juan², XIN Ming-Hong¹, QIU Shi-Lun¹. In-situ Synthesis and Activity of Nano-hydroxyapatite/Lysozyme Composite [J]. Chem. J. Chinese Universities, 2006, 27(8): 1421.
[13]	CHEN Kai, ZHAI Chun-Xi, LI Wen, WU Li-Xin, WU Yu-Qing, ZHANG Jian-Ping, AI Xi-Cheng . Time-resolved Fluorescence Spectroscopic Studies on the Binding of L-[4-(1-Pyrene)butyroyl]-phenylalanine with Proteins [J]. Chem. J. Chinese Universities, 2004, 25(10): 1905.
[14]	CHEN Yan-Tao, LUO Zhong-Lin, DING Jian-Dong . Preliminary Computer Simulation of Spatial Structure of SARS Coronavirus E-protein [J]. Chem. J. Chinese Universities, 2003, 24(8): 1406.
[15]	WANG Yan, LI Min, GONG Bo-Lin, GENG Xin-Du . Quantitative Characterization of Molecular Conformation of Lysozyme with Z in Liquid Chromatography [J]. Chem. J. Chinese Universities, 2003, 24(7): 1207.