Repeat regions R1 and R2 in the P97 paralogue Mhp271 of Mycoplasma hyopneumoniae bind heparin, fibronectin and porcine cilia

Deutscher, Ania T. and Jenkins, Cheryl and Minion, F. Chris and Seymour, Lisa M. and Padula, Matthew P. and Dixon, Nicholas E. and Walker, Mark J. and Djordjevic, Steven P. (2010) Repeat regions R1 and R2 in the P97 paralogue Mhp271 of Mycoplasma hyopneumoniae bind heparin, fibronectin and porcine cilia. Molecular Microbiology, 78 (2). pp. 444-458. ISSN 0950-382X

Abstract

Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, adheres to ciliated respiratory epithelia resulting in ciliostasis and epithelial cell death. The cilium adhesin P97 (Mhp183) contains two repeat regions, designated R1 and R2, that play key roles in adherence. Eight pentapeptide repeats in R1 are sufficient to bind porcine cilia; however, both R1 and R2 are needed to bind heparin. Mhp271, a paralogue of P97, is the only other M. hyopneumoniae protein to contain both R1 and R2 repeats. These repeats are arranged as a set of three pentapeptide repeats (designated R1A271), two decapeptide repeats (designated R2271), and a second set of six pentapeptide repeats (designated R1B271). To determine their function, recombinant proteins containing R1A271 (F1271) and R2271-R1B271 (F2271) were constructed and used in in vitro binding assays. F2271, but not F1271, bound heparin (KD = 8.1 ± 0.4 nM), fibronectin (KD = 174 ± 13 nM) and porcine cilia. Pre-incubation of F2271 with 100 μM heparin blocked cilium binding by ~69%. Cell surface shaving with trypsin combined with two-dimensional liquid chromatography coupled to tandem mass spectrometry analysis identified Mhp271 as surface-exposed. Our data suggest that both R1 and R2 in Mhp271 are involved in binding to host molecules.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: © 2010 Blackwell Publishing Ltd. Permanent restricted access to published version in accordance with the copyright policy of the publisher.
Faculty / Department / School: Historic - Faculty of Sciences - Department of Nursing
Date Deposited: 29 Jul 2014 02:03
Last Modified: 25 Aug 2014 06:23
Uncontrolled Keywords: adhesin; fibronectin; heparin; pentapeptide; recombinant protein; trypsin
Fields of Research : 07 Agricultural and Veterinary Sciences > 0702 Animal Production > 070205 Animal Protection (Pests and Pathogens)
06 Biological Sciences > 0606 Physiology > 060603 Animal Physiology - Systems
07 Agricultural and Veterinary Sciences > 0707 Veterinary Sciences > 070707 Veterinary Microbiology (excl. Virology)
Socio-Economic Objective: E Expanding Knowledge > 97 Expanding Knowledge > 970106 Expanding Knowledge in the Biological Sciences
Identification Number or DOI: 10.1111/j.1365-2958.2010.07345.x
URI: http://eprints.usq.edu.au/id/eprint/25086

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