hepatitis B;
L HBsAg;
phosphorylation;
NMR;
cell-free (CF) protein synthesis;
mass spectrometry;
SYNTHESIS SYSTEM;
DOMAIN;
IDENTIFICATION;
PURIFICATION;
INFECTION;
PRE-S1;
MODEL;
CORE;
D O I:
10.3389/fmolb.2021.821755
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
We here establish the phosphorylation sites in the human hepatitis B virus (HBV) large envelope protein (L). L is involved in several functionally important interactions in the viral life cycle, including with the HBV cellular receptor, HBV capsid, Hsc70 chaperone, and cellular membranes during fusion. We have recently shown that cell-free synthesis of the homologous L protein of duck HBV in wheat germ extract results in very similar phosphorylation events to those previously observed in animal cells. Here, we used mass spectrometry and NMR to establish the phosphorylation patterns of human HBV L protein produced by both in vitro cell-free synthesis and in E. coli with the co-expression of the human MAPK14 kinase. While in the avian virus the phosphorylation of L has been shown to be dispensable for infectivity, the identified locations in the human virus protein, both in the PreS1 and PreS2 domains, raise the intriguing possibility that they might play a functional role, since they are found at strategic sites predicted to be involved in L interactions. This would warrant the further investigation of a possible function in virion formation or cell entry.
机构:
Univ N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA
McKnight, Kevin L.
Xie, Ling
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机构:
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Xie, Ling
Gonzalez-Lopez, Olga
论文数: 0引用数: 0
h-index: 0
机构:
Univ N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Gonzalez-Lopez, Olga
Rivera-Serrano, Efrain E.
论文数: 0引用数: 0
h-index: 0
机构:
Univ N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Rivera-Serrano, Efrain E.
Chen, Xian
论文数: 0引用数: 0
h-index: 0
机构:
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Chen, Xian
Lemon, Stanley M.
论文数: 0引用数: 0
h-index: 0
机构:
Univ N Carolina, Dept Med, Chapel Hill, NC 27599 USA
Univ N Carolina, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA
Univ N Carolina, Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 USAUniv N Carolina, Dept Med, Chapel Hill, NC 27599 USA