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P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae : defined domains of P159 bind heparin and promote adherence to eukaryote cells
Author(s) -
Burnett Tracey A.,
Dinkla Katrin,
Rohde Manfred,
Chhatwal Gursharan S.,
Uphoff Cord,
Srivastava Mukesh,
Cordwell Stuart J.,
Geary Steven,
Liao Xiaofen,
Minion F. Chris,
Walker Mark J.,
Djordjevic Steven P.
Publication year - 2006
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2006.05139.x
Subject(s) - mycoplasma hyopneumoniae , bacterial adhesin , biology , heparin , microbiology and biotechnology , recombinant dna , secretion , eukaryote , biochemistry , escherichia coli , gene , genome
Summary Mycoplasma hyopneumoniae , the causative agent of porcine enzootic pneumonia, colonizes the respiratory cilia of affected swine causing significant economic losses to swine production worldwide. Heparin is known to inhibit adherence of M. hyopneumoniae to porcine respiratory epithelial cilia. M. hyopneumoniae cells bind heparin but the identity of the heparin‐binding proteins is limited. Proteomic analysis of M. hyopneumoniae lysates identified 27 kDa (P27), 110 kDa (P110) and 52 kDa (P52) proteins representing different regions of a 159 kDa (P159) protein derived from mhp494. These cleavage fragments were surface located and present at all growth stages. Following purification of four recombinant proteins spanning P159 (F1 P159 , F2 P159 , F3 P159 and F4 P159 ), only F3 P159 and F4 P159 bound heparin in a dose‐dependent manner ( K d values 142.37 ± 22.01 nM; 75.37 ± 7.34 nM respectively). Scanning electron microscopic studies showed M. hyopneumoniae bound intimately to porcine kidney epithelial‐like cells (PK15 cells) but these processes were inhibited by excess heparin and F4 P159 . Similarly, latex beads coated with F2 P159 and F4 P159 adhered to and entered PK15 cells, but heparin, F2 P159 and F4 P159 was inhibitory. These findings indicate that P159 is a post‐translationally cleaved, glycosaminoglycan‐binding adhesin of M. hyopneumoniae .