Bioinformatics analysis on structure and function and expression vector construction of SARS-CoV-2 related protein TMPRSS2
- Author:
Ben-Jin XU
1
;
Hong-Rong YAN
1
;
Miao DU
1
;
Yan XUAN
1
;
Yan-Xiang HOU
1
;
Ya-Nan YANG
1
;
Ling LIU
1
;
Zhuo-Xi LI
2
;
Lei FAN
2
;
Jing LI
2
;
Xiao-Cong CHEN
2
;
Wen-Ting TANG
2
;
Jun-Xiao YU
2
;
Jie MEN
3
;
Bin-Yu SONG
4
;
Xiao-Liang LIU
4
Author Information
- Publication Type:Journal Article
- Keywords: bioinformatics analysis; evolutionary' analysis; SARS-CoV-2; structure and function; TMPRSS2; vector construction
- From: Chinese Pharmacological Bulletin 2022;38(8):1218-1226
- CountryChina
- Language:Chinese
- Abstract: Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.
