Identification of a region in the S1 subunit of pertussis toxin that is required for enzymatic activity and that contributes to the formation of a neutralizing antigenic determinant.
The S1 subunit of pertussis toxin possesses two regions (homology boxes), each spanning 8 residues, that are nearly identical in sequence to similarly located regions in the enzymatically active A fragments of two other ADP-ribosylating toxins: cholera toxin and Escherichia coli heat-labile toxin. This observation suggests a functional role for one or both of these regions in enzymatic activity. We have examined the role of one of these regions, located near the amino terminus of the S1 subunit, by using a high-level recombinant expression system and progressive truncation of the gene sequence encoding the amino terminus of the molecule. A series of six truncated, recombinant proteins were produced at high levels in E. coli and examined for their enzymatic and antigenic properties. The three molecules that lacked most or all of the homology box delimited by amino acid residues 8 and 15 lacked detectable enzymatic activity. All of the three molecules in which the box was retained exhibited detectable activity. Only those recombinant molecules that possessed the homology box reacted with a neutralizing and passively protective monoclonal anti-S1 antibody. These findings identify the region of homology located near the amino terminus of S1 as an apparent enzymatic subsite and a potentially important antigenic determinant.
Cieplak
W
W
Department of Health and Human Services, National Institute of Allergy and Infectious Diseases, Hamilton, MT 59840.
Burnette
W N
WN
Mar
V L
VL
Kaljot
K T
KT
Morris
C F
CF
Chen
K K
KK
Sato
H
H
Keith
J M
JM
D000246:Adenosine Diphosphate Ribose / Q000096:biosynthesis; D000595:Amino Acid Sequence; D000939:Epitopes / Q000276:immunology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D037342:Pertussis Toxin; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000939:Epitopes; D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D037342:Pertussis Toxin