Abstract
To identify the major antigenic determinant of native Salmonella flagella of antigenic type d, we constructed a series of mutated fliC(d) genes with deletions and amino acid alterations in hypervariable region IV and in regions of putative epitopes as suggested by epitope mapping with synthetic octameric peptides (T.M. Joys and F. Schodel, Infect. Immun. 59:3330-3332, 1991). The expressed product of most of the mutant genes, with deletions of up to 92 amino acids in region IV, assembled into functional flagella and conferred motility on flagellin-deficient hosts. Serological analysis of these flagella with different anti-d antibodies revealed that the peptide sequence centered at amino acids 229 to 230 of flagellin was a dominant B- cell epitope at the surface of d flagella, because replacement of these two amino acids alone or together with their flanking sequence by a tripeptide specified by a linker sequence eliminated most reactivity with antisera against wild-type d flagella as tested by enzyme-linked immunosorbent assay or by Western immunoblot. Functional analysis of the mutated flagella genes with or without an insert suggested that amino acids 180 to 214 in the 5' part of hypervariable region IV (residues 181 to 307 of the total of 505) is important to the function of flagella. The hybrid proteins formed by insertion of peptide sequence pre-S1 12-47 of hepatitis B virus surface antigen into the deleted flagellins assembled into functional flagella, and antibody to the pre-S1 sequence was detected after immunization of mice with the hybrid protein. This suggests that such mutant flagellins containing heterologous epitopes have potential as vaccines.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2406-2414 |
| Number of pages | 9 |
| Journal | Journal of Bacteriology |
| Volume | 176 |
| Issue number | 8 |
| State | Published - 1994 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Immunology
Cite this
Hypervariable region IV of Salmonella gene fliC(d) encodes a dominant surface epitope and a stabilizing factor for functional flagella. / He, Xiaosong; Rivkina, M.; Stocker, B. A D; Robinson, W. S.
In: Journal of Bacteriology, Vol. 176, No. 8, 1994, p. 2406-2414.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Hypervariable region IV of Salmonella gene fliC(d) encodes a dominant surface epitope and a stabilizing factor for functional flagella
AU - He, Xiaosong
AU - Rivkina, M.
AU - Stocker, B. A D
AU - Robinson, W. S.
PY - 1994
Y1 - 1994
N2 - To identify the major antigenic determinant of native Salmonella flagella of antigenic type d, we constructed a series of mutated fliC(d) genes with deletions and amino acid alterations in hypervariable region IV and in regions of putative epitopes as suggested by epitope mapping with synthetic octameric peptides (T.M. Joys and F. Schodel, Infect. Immun. 59:3330-3332, 1991). The expressed product of most of the mutant genes, with deletions of up to 92 amino acids in region IV, assembled into functional flagella and conferred motility on flagellin-deficient hosts. Serological analysis of these flagella with different anti-d antibodies revealed that the peptide sequence centered at amino acids 229 to 230 of flagellin was a dominant B- cell epitope at the surface of d flagella, because replacement of these two amino acids alone or together with their flanking sequence by a tripeptide specified by a linker sequence eliminated most reactivity with antisera against wild-type d flagella as tested by enzyme-linked immunosorbent assay or by Western immunoblot. Functional analysis of the mutated flagella genes with or without an insert suggested that amino acids 180 to 214 in the 5' part of hypervariable region IV (residues 181 to 307 of the total of 505) is important to the function of flagella. The hybrid proteins formed by insertion of peptide sequence pre-S1 12-47 of hepatitis B virus surface antigen into the deleted flagellins assembled into functional flagella, and antibody to the pre-S1 sequence was detected after immunization of mice with the hybrid protein. This suggests that such mutant flagellins containing heterologous epitopes have potential as vaccines.
AB - To identify the major antigenic determinant of native Salmonella flagella of antigenic type d, we constructed a series of mutated fliC(d) genes with deletions and amino acid alterations in hypervariable region IV and in regions of putative epitopes as suggested by epitope mapping with synthetic octameric peptides (T.M. Joys and F. Schodel, Infect. Immun. 59:3330-3332, 1991). The expressed product of most of the mutant genes, with deletions of up to 92 amino acids in region IV, assembled into functional flagella and conferred motility on flagellin-deficient hosts. Serological analysis of these flagella with different anti-d antibodies revealed that the peptide sequence centered at amino acids 229 to 230 of flagellin was a dominant B- cell epitope at the surface of d flagella, because replacement of these two amino acids alone or together with their flanking sequence by a tripeptide specified by a linker sequence eliminated most reactivity with antisera against wild-type d flagella as tested by enzyme-linked immunosorbent assay or by Western immunoblot. Functional analysis of the mutated flagella genes with or without an insert suggested that amino acids 180 to 214 in the 5' part of hypervariable region IV (residues 181 to 307 of the total of 505) is important to the function of flagella. The hybrid proteins formed by insertion of peptide sequence pre-S1 12-47 of hepatitis B virus surface antigen into the deleted flagellins assembled into functional flagella, and antibody to the pre-S1 sequence was detected after immunization of mice with the hybrid protein. This suggests that such mutant flagellins containing heterologous epitopes have potential as vaccines.
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M3 - Article
C2 - 7512552
AN - SCOPUS:0028226746
VL - 176
SP - 2406
EP - 2414
JO - Journal of Bacteriology
JF - Journal of Bacteriology
SN - 0021-9193
IS - 8
ER -