[Conserved amino-acid residue mutations in epitope of human cytomegalovirus antigen M].
We identified the critical amino-acid residues in antigen M derterminant (MAD) epitope of human cytomegalovirus protein M. On the basis of the peptide sequence of MAD, some conservative residues were mutated into the glycine residue. Then the gene fragment of mutants linked to amino terminal of Fc were cloned into the plasmid pET32-Fc and expressed by fusion with Fc. After purified by protein A affinity chromatography, the activity of mutants binding the goat polyclonal antibodies against human cytomegalovirus (HCMV) were detected by ELISA and Western blotting. Our results showed that when glutamine residue was mutated into glycine residue, the activity of MAD(Q --> G) binding the goat polyclonal antibodies against HCMV was reduced apparently. Other mutants did not have the same characteristics. The activity of MAD was closely related to the conformation of glutamine residue.
Wang
Benxu
B
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Liu
Zhan
Z
Liu
Yu
Y
Shen
Beifen
B
Liu
Chuan
C
Shao
Ningsheng
N
D000596:Amino Acids / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D003587:Cytomegalovirus / Q000235:genetics* / Q000276:immunology; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D006041:Goats; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000235:genetics; D009154:Mutation; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000276:immunology
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D007141:Immunoglobulin Fc Fragments; D050505:Mutant Proteins; D014763:Viral Matrix Proteins