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Infection and Immunity, March 2005, p. 1515-1522, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1515-1522.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Effect of Mutations in the Human Immunoglobulin A1 (IgA1) Hinge on Its Susceptibility to Cleavage by Diverse Bacterial IgA1 Proteases

Bernard W. Senior¹ and Jenny M. Woof^1*

Division of Pathology and Neuroscience, University of Dundee Medical School, Dundee, United Kingdom¹

Received 1 October 2004/ Returned for modification 10 November 2004/ Accepted 15 November 2004

Components of the human immunoglobulin A1 (IgA1) hinge governing sensitivity to cleavage by bacterial IgA1 proteases were investigated. Recombinant antibodies with distinct hinge mutations were constructed from a hybrid comprised of human IgA2 bearing half of the human IgA1 hinge region. This hybrid antibody and all the mutant antibodies derived from it were resistant to cleavage by the IgA1 proteases from Streptococcus oralis and Streptococcus mitis biovar 1 strains but were cleaved to various degrees by those of Streptococcus pneumoniae, some Streptococcus sanguis strains, and the type 1 and 2 IgA1 proteases of Haemophilus influenzae, Neisseria meningitidis, and Neisseria gonorrhoeae. Remarkably, those proteases that cleave a Pro-Ser peptide bond in the wild-type IgA1 hinge were able to cleave mutant antibodies lacking a Pro-Ser peptide bond in the hinge, and those that cleave a Pro-Thr peptide bond in the wild-type IgA1 hinge were able to cleave mutant antibodies devoid of a Pro-Thr peptide bond in the hinge. Thus, the enzymes can cleave alternatives to their preferred postproline peptide bond when such a bond is unavailable. Peptide sequence analysis of a representative antibody digestion product confirmed this conclusion. The presence of a cleavable peptide bond near the CH2 end of the hinge appeared to result in greater cleavage than if the scissile bond was at the CH1 end of the hinge. Proline-to-serine substitution at residue 230 in a hinge containing potentially cleavable Pro-Ser and Pro-Thr peptide bonds increased the resistance of the antibody to cleavage by many IgA1 proteases.

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* Corresponding author. Mailing address: Division of Pathology and Neuroscience, University of Dundee Medical School, Ninewells Hospital, Dundee, United Kingdom. Phone: 44 1382 660111, ext. 33540. Fax: 44 1382 633952. E-mail: j.m.woof{at}dundee.ac.uk.

Editor: D. L. Burns

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Infection and Immunity, March 2005, p. 1515-1522, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1515-1522.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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