Mapping the intersubunit region of influenza virus hemagglutinin: comparative CD and FTIR spectroscopic studies on multiple antigenic peptides.
An A/PR/8/34 (IHN1) influenza virus hemagglutinin (HA)-specific monoclonal antibody (Z38) was found to react with the solid phase adsorbed influenza virus expressing uncleaved (HA0) molecules but not to bind to virus particles bearing enzymatically cleaved hemagglutinin. Synthetic peptides corresponding to the uncleaved HA0 317-341 intersubunit region of subtypes H1-H3 (IP1-IP3) or comprising either the C-terminal 317-329 amino acids of HA1 (CP1) or the N-terminal 330-341 of HA2 (fusion peptide, FP) subunits of cleaved HA were used to characterize the fine specificity of Z38 mAb. Circular dichroism and Fourier-transform infrared spectroscopy showed that, compared to IP2 and IP3 comprising the H2 and H3 subtype fragments of the intact intersubunit region, IP1 has relatively low helicity but a tendency to adopt beta-turns in trifluoroethanol. The immunological and conformational properties of multiple antigenic peptides (MAPs) containing four copies of CP1 were also studied. Based on the appearance of an infrared component band at 1637 cm-1 (beta-turn band), the CP1 arms of MAPs also contain repeats of beta-turns. However, it is only the MAP1-FP construct comprising also the fusion peptide, which binds Z38 mAb as strongly as IP1 does. This puts emphasis on the role of the fusion region in modifying conformation and consequently the ability of peptides to elicit an antibody response. The results obtained for peptide conformation also suggests a beta-turn(s)/beta-sheet/beta-turn/beta-sheet conformational motif in the recognition by the hemagglutinin subtype-specific Z38 monoclonal antibody or by peptide-induced polyclonal antibodies.
Majer
Z
Z
Department of Organic Chemistry, Eötvös University, Budapest, Hungary.
Holly
S
S
Tóth
G K
GK
Váradi
G
G
Nagy
Z
Z
Horváth
A
A
Rajnavölgyi
E
E
Laczkó
I
I
Hollósi
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002942:Circular Dichroism; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000235:genetics*; D009980:Influenza A virus / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017550:Spectroscopy, Fourier Transform Infrared
D000911:Antibodies, Monoclonal; D006389:Hemagglutinins, Viral; D010455:Peptides; D011993:Recombinant Fusion Proteins