Poster Presentation Lorne Infection and Immunity 2019

Shark antibodies and their role in targeting bacterial adhesins: alternatives against urinary tract infections (#156)

Gabriela Constanza Martinez Ortiz 1 , Jason Paxman 1 , Julieanne Vo 1 , Geqing Wang 1 , Kevin Lim 1 , Katherine Griffiths 1 , Michael Foley 1 , Mark Schembri 2 , Begoña Heras 1
  1. Department of Biochemistry, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC 3086, Australia
  2. Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane , QLD 4072, Australia

Urinary tract infections (UTIs) are one of the most common bacterial infections in humans [4]. Treatments for these infections are becoming scarcer due to the increasing bacterial antibiotic resistance [6]. Therefore, new approaches to tackle UTIs are urgently required. The main cause of UTIs is Uropathogenic Escherichia coli (UPEC), a pathogen that encodes multiple virulence factors, which play a key role in the establishment and persistence of the infection [3, 4, 5]. The biggest family of secreted virulent proteins in E. coli is the Autotransporter proteins (ATs) family, which contains an important group known as the AIDA-I-type adhesins [1, 2, 8]. These surface proteins promote multiple phenotypes including  bacterial aggregation and biofilm formation [2, 6, 8]. This study is focused on the most prevalent E. coli AT adhesin, Antigen 43 (Ag43) [7]. Specifically, this research aims at analysing structure-function relationships, processing and mechanism of action of Ag43, in order to develop an approach to block its function. The functional domain of Ag43b  from  the UPEC  strain CFT073 was purified and the structure was characterised by X-ray crystallography.

The identification of molecules that block the function of Ag43 has been attempted using single-domain antibodies derived from the variable region (vNAR) of a shark antibody known as the IgNAR. Screening of a vNAR phage-displayed library resulted in the identification of two binders that specifically recognise Ag43 homologues. Ongoing work with these vNARs will establish their ability to halt the aggregative ability of Ag43. 

The outcomes of this project will provide essential data for future studies on the function and inhibition of this key AT to allow the future development of alternative therapeutic approaches to tackle UTIs.

  1. [1] Celik N, Webb CT, Leyton DL, Holt KE, Heinz E, Gorrell R, Kwok T, Naderer T, Strugnell RA, Speed TP, Teasdale RD, Likic VA, Lithgow T (2012) A bioinformatic strategy for the detection, classification and analysis of bacterial autotransporters, PloS one, 7: e43245
  2. [2] Henderson IR, Nataro JP (2001) Virulence functions of autotransporter proteins, Infect Immun, 69: 1231-1243
  3. [3] Lüthje P, Brauner A (2010) Ag43 promotes persistence of Uropathogenic Escherichia coli in the urinary tract, J Clin Microbiol, 48(6): 2316–2317
  4. [4] Mulvey MA, Schilling JD, Hultgreen SC (2001) Establishment of a persistent Escherichia coli reservoir during the acute phase of a bladder infection Infect Immun, 69(7): 4572–4579
  5. [5] Schreiber HL 4th, Conover MS, Chou WC, Hibbing ME, Manson AL, Dodson KW, Hannan TJ, Roberts PL, Stapleton AE, Hooton TM, Livny J, Earl AM, Hultgren SJ (2017) Bacterial virulence phenotypes of Escherichia coli and host susceptibility determine risk for urinary tract infections, Sci Transl Med, 9(382). pii: eaaf1283
  6. [6] van Ulsen P, Rahman Su, Jong WSP, Daleke-Schermerhorn MH, Luirink J (2013) Type V secretion: From biogenesis to biotechnology, Biochim Biophys Acta, 1843(8): 1592-611
  7. [7] Vo JL, Martinez-Ortiz GC, Subedi P, Keerthikumar S, Mathivanan S, Paxman J, Heras B (2017) Autotransporter Adhesins in Escherichia coli Pathogenesis, Proteomics, 17: 1600431-n/a
  8. [8] Wells TJ, Totsika M, Schembri MA (2010) Autotransporters of Escherichia coli: a sequence based characterization, Microbiology, 156: 2459-2469