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:
Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Mehmankhah, Mahboubeh
Bhat, Ruchika
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Indian Inst Technol Delhi, Dept Chem & Sch Biol Sci, New Delhi 110016, India
Indian Inst Technol Delhi, Supercomp Facil Bioinformat & Computat Biol SCF, New Delhi 110016, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Bhat, Ruchika
Anvar, Mohammad Sabery
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Natl Inst Genet Engn & Biotechnol, Syst Biotechnol Dept, Tehran, IranJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Anvar, Mohammad Sabery
Ali, Shahnawaz
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Ali, Shahnawaz
Alam, Aftab
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Alam, Aftab
Farooqui, Anam
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Farooqui, Anam
Amir, Fatima
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Amir, Fatima
Anwer, Ayesha
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Anwer, Ayesha
Khan, Saniya
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Khan, Saniya
Azmi, Iqbal
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Jamia Millia Islamia, Multidisciplinary Ctr Adv Res & Studies, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Azmi, Iqbal
Ali, Rafat
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Ali, Rafat
Ishrat, Romana
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Ishrat, Romana
Hassan, Imtaiyaz
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Hassan, Imtaiyaz
Minuchehr, Zarrin
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Natl Inst Genet Engn & Biotechnol, Syst Biotechnol Dept, Tehran, IranJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India
Minuchehr, Zarrin
Kazim, Syed Naqui
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Jamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, IndiaJamia Millia Islamia, Ctr Interdisciplinary Res Basic Sci, New Delhi 110025, India