Concomitant loss of conformation and superantigenic activity of staphylococcal enterotoxin B deletion mutant proteins.

Abstract:

:The T-cell-stimulating activity of staphylococcal enterotoxin B (SEB) is an important factor in the pathogenesis of certain staphylococcal diseases. To investigate the immunologically active domains of the SEB molecule, we have produced truncated fragments of recombinant SEB by C-terminal and N-terminal deletions. The fragments were expressed as fusion proteins with protein A, including a cleavage site to remove the protein A part. Mutant proteins were tested for the ability to stimulate human resting T cells and SEB-reactive T-cell clones. Deletion of only 9 amino acids from the C terminus leads to complete loss of T-cell-stimulating activity. Removing further amino acids from the SEB molecule did not lead to a reexpression of T-cell-mitogenic activity. A mutant protein, however, in which the 9 C-terminal amino acids were replaced with a tail of 68 amino acids encoded by the vector was fully active. Two mutant proteins with N-terminal deletions of 60 and 81 amino acids were inactive as well. A neutralizing monoclonal antibody against a conformational epitope lost binding with all the inactive mutant proteins only, whereas a monoclonal antibody recognizing an epitope involved in emetic activity reacted with all mutant proteins. These results suggest that even small deletions at the C terminus affect the three-dimensional conformation of the SEB molecule.

journal_name

Infect Immun

journal_title

Infection and immunity

authors

Metzroth B,Marx T,Linnig M,Fleischer B

doi

10.1128/IAI.61.6.2445-2452.1993

subject

Has Abstract

pub_date

1993-06-01 00:00:00

pages

2445-52

issue

6

eissn

0019-9567

issn

1098-5522

journal_volume

61

pub_type

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