APM OpenIR
Recombinant Butelase-Mediated Cyclization of the p53-Binding Domain of the Oncoprotein MdmX-Stabilized Protein Conformation as a Promising Model for Structural Investigation
Pi, Ni1,2; Gao, Meng1,2; Cheng, Xiyao1,2,3; Liu, Huili4; Kuang, Zhengkun1,2; Yang, Zixin1,2; Yang, Jing1,2; Zhang, Bailing1,2; Chen, Yao1,2; Liu, Sen1,2; Huang, Yongqi1,2,3; Su, Zhengding1,2,3
2019-07-09
Source PublicationBIOCHEMISTRY
ISSN0006-2960
Volume58Issue:27Pages:3005-3015
AbstractCyclization of the polypeptide backbone has proven to be a powerful strategy for enhancing protein stability for fundamental research and pharmaceutical application. The use of such an approach is restricted by how well a targeted polypeptide can be efficiently ligated. Recently, an Asx-specific peptide ligase identified from a tropical cyclotide-producing plant and named butelase 1 exhibited excellent cyclization kinetics that cannot be matched by other known ligases, including intein, PATG, PCY1, and sortase A. In this work, we aimed to examine whether butelase 1 facilitated conformational stability for structural investigation First we successfullly expressed recombined butelase 1 (rBTase) in the yeast Pichia pastoris. Next, rBTase was shown to be highly efficient in the cyclization of the p53-binding domain (N-terminal domain) of murine double minute X (N-MdmX), an important target for designing anticancer drugs. The cyclized N-MdmX (cMdmX) exhibited increased conformational stability and improved interaction with the ligand compared with those of noncyclized N-MdmX. Importantly, the thermal melting process was completely reversible, contrary to noncyclized N-MdmX, and the melting temperature (T-m) of cMdmX was increased to 47 from 43 degrees C. This stable conformation of cMdmX was further confirmed by N-15-H-1 heteronuclear single-quantum coherence nuclear magnetic resonance (NMR) spectroscopy. The complex of cMdmX and the ligand was tested for protein crystallization, and several promising findings were revealed. Therefore, our work not only provides a recombinant version of butelase 1 but also suggests a conventional approach for preparing stable protein samples for both protein crystallization and NMR structural investigation.
Funding OrganizationNational Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China
DOI10.1021/acs.biochem.9b00263
WOS KeywordPEPTIDES ; MACROCYCLIZATION ; INHIBITOR ; LIGATION ; BINDING ; P53
Language英语
Funding ProjectNational Natural Science Foundation of China[21603121] ; National Natural Science Foundation of China[3167076831500132] ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics[T152604] ; Wuhan Science and Technology Bureau of China[2018060401011319]
Funding OrganizationNational Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics ; Wuhan Science and Technology Bureau of China ; Wuhan Science and Technology Bureau of China
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemistry & Molecular Biology
WOS IDWOS:000475408700006
PublisherAMER CHEMICAL SOC
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.apm.ac.cn/handle/112942/14644
Collection中国科学院武汉物理与数学研究所
Corresponding AuthorHuang, Yongqi; Su, Zhengding
Affiliation1.Hubei Univ Technol, Hubei Key Lab Ind Microbiol, Minist Educ, Key Lab Ind Fermentat, Wuhan 430068, Hubei, Peoples R China
2.Hubei Univ Technol, Natl Ctr Cellular Regulat & Mol Pharmaceut 111, Wuhan 430068, Hubei, Peoples R China
3.Wuhan Amersino Biodevelop Inc, B1 Bldg,Biolake Pk, Wuhan 430075, Hubei, Peoples R China
4.Chinese Acad Sci, Wuhan Inst Phys & Math, State Key Lab Magnet Resonance & Atom & Mol Phys, Natl Ctr Magnet Resonance Wuhan, Wuhan 430071, Hubei, Peoples R China
Recommended Citation
GB/T 7714
Pi, Ni,Gao, Meng,Cheng, Xiyao,et al. Recombinant Butelase-Mediated Cyclization of the p53-Binding Domain of the Oncoprotein MdmX-Stabilized Protein Conformation as a Promising Model for Structural Investigation[J]. BIOCHEMISTRY,2019,58(27):3005-3015.
APA Pi, Ni.,Gao, Meng.,Cheng, Xiyao.,Liu, Huili.,Kuang, Zhengkun.,...&Su, Zhengding.(2019).Recombinant Butelase-Mediated Cyclization of the p53-Binding Domain of the Oncoprotein MdmX-Stabilized Protein Conformation as a Promising Model for Structural Investigation.BIOCHEMISTRY,58(27),3005-3015.
MLA Pi, Ni,et al."Recombinant Butelase-Mediated Cyclization of the p53-Binding Domain of the Oncoprotein MdmX-Stabilized Protein Conformation as a Promising Model for Structural Investigation".BIOCHEMISTRY 58.27(2019):3005-3015.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Pi, Ni]'s Articles
[Gao, Meng]'s Articles
[Cheng, Xiyao]'s Articles
Baidu academic
Similar articles in Baidu academic
[Pi, Ni]'s Articles
[Gao, Meng]'s Articles
[Cheng, Xiyao]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Pi, Ni]'s Articles
[Gao, Meng]'s Articles
[Cheng, Xiyao]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.