P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae: defined domains of P159 bind heparin and promote adherence to eukaryote cells.
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 (F1P159, F2P159, F3P159 and F4P159), only F3P159 and F4P159 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 F4P159. Similarly, latex beads coated with F2P159 and F4P159 adhered to and entered PK15 cells, but heparin, F2P159 and F4P159 was inhibitory. These findings indicate that P159 is a post-translationally cleaved, glycosaminoglycan-binding adhesin of M. hyopneumoniae.
Burnett
Tracey A
TA
NSW Department of Primary Industries, Elizabeth Macarthur Agricultural Institute, Camden, NSW 2570, Australia.
Dinkla
Katrin
K
Rohde
Manfred
M
Chhatwal
Gursharan S
GS
Uphoff
Cord
C
Srivastava
Mukesh
M
Cordwell
Stuart J
SJ
Geary
Steven
S
Liao
Xiaofen
X
Minion
F Chris
FC
Walker
Mark J
MJ
Djordjevic
Steven P
SP
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion; D002460:Cell Line; D004847:Epithelial Cells / Q000382:microbiology; D006493:Heparin / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000382:microbiology*; D008969:Molecular Sequence Data; D045705:Mycoplasma hyopneumoniae / Q000378:metabolism* / Q000502:physiology; D045729:Pneumonia of Swine, Mycoplasmal / Q000382:microbiology; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013552:Swine
D018829:Adhesins, Bacterial; D011994:Recombinant Proteins; D006493:Heparin