1887

Abstract

Major poultry pathogens and share a gene encoding a putative cysteine protease CysP similar to papain cysteine protease (C1A subfamily). Comparison of the gene sequences of 18 and 10 strains sequenced in this study showed polymorphisms, including deletions. Seven strains, including the type strain WVU 1853, had a 39 bp deletion in the 3′ end of the gene. In the same region, all strains showed a deletion of 66 bp. Immunoblot analysis with specific antibodies demonstrated that strains expressed CysP, which was approximately 65 kDa. Both and were able to digest chicken IgG (cIgG). Incubation of cIgG (∼170 kDa) with or cells (∼15 h at 37 °C) resulted in a papain-like cleavage pattern of cIgG and fragments corresponding to the antigen-binding fragment of IgG (Fab, ∼45 kDa) and the crystallizable region fragment (Fc) of the IgG heavy chain (dimer of ∼60 kDa). Iodoacetamide (50 mM) prevented cleavage of cIgG by both species. Following site-directed mutagenesis (eight TGA codons were changed to TGG) the gene of ULB 925 was expressed as a His-tagged protein in a cell-free system. Purified recombinant CysP (rCysP; ∼67 kDa, pI∼8) cleaved cIgG into Fab and Fc fragments. This indicates that CysP is responsible for the cIgG cleavage caused by and, probably, by . This is the first evidence to our knowledge that mycoplasmas have enzymes that can cleave the host IgG and indicates a novel strategy used by and for prolonged survival despite the antibody response of their host.

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2011-02-01
2020-01-17
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