The antigen 43 structure reveals a molecular Velcro-like mechanism of autotransporter-mediated bacterial clumping
Open accessAbstract
Aggregation and biofilm formation are critical mechanisms for bacterial resistance to host immune factors and antibiotics. Autotransporter (AT) proteins, which represent the largest group of outer-membrane and secreted proteins in Gram-negative bacteria, contribute significantly to these phenotypes. Despite their abundance and role in bacterial pathogenesis, most AT proteins have not been structurally characterized, and there is a paucity of detailed information with regard to their mode of action. Here we report the structure-function relationships of Antigen 43 (Ag43a), a prototypic self-associating AT protein from uropathogenic Escherichia coli. The functional domain of Ag43a displays a twisted L-shaped β-helical structure firmly stabilized by a 3D hydrogen-bonded scaffold. Notably, the distinctive Ag43a L shape facilitates self-association and cell aggregation. Combining all our data, we define a molecular "Velcro-like" mechanism of AT-mediated bacterial clumping, which can be tailored to fit different bacterial lifestyles such as the formation of biofilms.
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The L-shape of Ag43 is important for autoaggregation, as mutations straightening the β-helical spine of the protein abolished the ability to autoaggregate.
“The L-shape of Ag43 is also important for autoaggregation, as mutations straightening the β-helical spine of the protein abolished the ability to autoaggregate [77].”
From Bacterial autoaggregation (2018) · cites this paper as [77]
Antigen 43 (Ag43) from E. coli self-associates through a polar interaction network, with both hydrogen bonds and salt bridges.
“In contrast to Hap, Antigen 43 (Ag43) from E. coli self-associates through a polar interaction network, with both hydrogen bonds and salt bridges [77].”
From Bacterial autoaggregation (2018) · cites this paper as [77]
The Ag43 extracellular region forms an L-shaped β-helix, with the self-association interface in the stem of the L in a ladder-like configuration of interacting residues reminiscent of Hap, and the interaction is in a trans orientation.
“The Ag43 extracellular region forms an L-shaped β-helix (Figure 2B). The self-association interface resides in the “stem” of the L, with a ladder-like configuration of interacting residues reminiscent of Hap. Also like Hap, the interaction is in a trans orientation.”
From Bacterial autoaggregation (2018) · cites this paper as [77]
In some cases, such as Ag43, a number of point mutations are required to prevent autoaggregation.
“In some cases, such as Ag43, a number of point mutations are required to prevent autoaggregation [77].”
From Bacterial autoaggregation (2018) · cites this paper as [77]
Cites
[20] Processing of X-ray diffraction data collected in oscillation mode
Otwinowski, Z., & Minor, W. (1997) · Methods in Enzymology · doi:10.1016/s0076-6879(97)76066-x · Cited by 33,475 · see citing works
<i>Coot</i> : model-building tools for molecular graphics
Emsley, P., & Cowtan, K. (2004) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444904019158 · Cited by 30,571 · see citing works
Kabsch, W. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909047337 · Cited by 16,463 · see citing works
NMRPipe: A multidimensional spectral processing system based on UNIX pipes
Delaglio, F., Grzesiek, S., Vuister, G. W., Zhu, G., Pfeifer, J., & Bax, A. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00197809 · Cited by 15,994 · see citing works
Guzman, L., Belin, D., Carson, M. J., & Beckwith, J. (1995) · Journal of Bacteriology · doi:10.1128/jb.177.14.4121-4130.1995 · Cited by 4,942 · see citing works
Scaling and assessment of data quality
Evans, P. (2005) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444905036693 · Cited by 4,391 · see citing works
<i>PHENIX</i> : building new software for automated crystallographic structure determination
Adams, P. D., Grosse-Kunstleve, R. W., Hung, L.-W., Ioerger, T. R., McCoy, A. J., Moriarty, N. W., Read, R. J., Sacchettini, J. C., Sauter, N. K., & Terwilliger, T. C. (2002) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444902016657 · Cited by 4,361 · see citing works
Optimal description of a protein structure in terms of multiple groups undergoing TLS motion
Painter, J., & Merritt, E. A. (2006) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444906005270 · Cited by 1,244 · see citing works
Intracellular Bacterial Biofilm-Like Pods in Urinary Tract Infections
Anderson, G. G., Palermo, J. J., Schilling, J. D., Roth, R., Heuser, J., & Hultgren, S. J. (2003) · Science · doi:10.1126/science.1084550 · Cited by 1,079 · see citing works
Uncomplicated Urinary Tract Infection
Hooton, T. M. (2012) · New England Journal of Medicine · doi:10.1056/nejmcp1104429 · Cited by 890 · see citing works
Type V Protein Secretion Pathway: the Autotransporter Story
Henderson, I. R., Navarro‐García, F., Desvaux, M., Fernandez, R. C., & Ala’Aldeen, D. A. A. (2004) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.68.4.692-744.2004 · Cited by 839 · see citing works
Serruto, D., Bottomley, M. J., Ram, S., Giuliani, M. M., & Rappuoli, R. (2012) · Vaccine · doi:10.1016/j.vaccine.2012.01.033 · Cited by 405 · see citing works
Structure of the translocator domain of a bacterial autotransporter
Oomen, C. J., van Ulsen, P., Gelder, P. V., et al. (2004) · The EMBO Journal · doi:10.1038/sj.emboj.7600148 · Cited by 363 · see citing works
Development and maturation of Escherichia coli K‐12 biofilms
Reisner, A., Haagensen, J. A. J., Schembri, M. A., Zechner, E. L., & Molin, S. (2003) · Molecular Microbiology · doi:10.1046/j.1365-2958.2003.03490.x · Cited by 352 · see citing works
Self-citationDanese, P. N., Pratt, L. A., Dove, S. L., & Kolter, R. (2000) · Molecular Microbiology · doi:10.1046/j.1365-2958.2000.02008.x · Cited by 345 · see citing works
Structure of Bordetella pertussis virulence factor P.69 pertactin
Emsley, P., Charles, I. G., Fairweather, N. F., & Isaacs, N. W. (1996) · Nature · doi:10.1038/381090a0 · Cited by 318 · see citing works
Discovery of an archetypal protein transport system in bacterial outer membranes
Selkrig, J., Mosbahi, K., Webb, C. T., Belousoff, M. J., Perry, A., Wells, T. J., Morris, F. C., Leyton, D. L., Totsika, M., Phan, M., et al. (2012) · Nature Structural & Molecular Biology · doi:10.1038/nsmb.2261 · Cited by 223 · see citing works
Self-citationFrom self sufficiency to dependence: mechanisms and factors important for autotransporter biogenesis
Leyton, D. L., Rossiter, A. E., & Henderson, I. R. (2012) · Nature Reviews Microbiology · doi:10.1038/nrmicro2733 · Cited by 216 · see citing works
Ulett, G. C., Valle, J., Beloin, C., Sherlock, O., Ghigo, J.-M., & Schembri, M. A. (2007) · Infection and Immunity · doi:10.1128/iai.01952-06 · Cited by 214 · see citing works
Self-citationIeva, R., & Bernstein, H. D. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0907912106 · Cited by 186 · see citing works
Otto, B. R., Sijbrandi, R., Luirink, J., Oudega, B., Heddle, J. G., Mizutani, K., Park, S., & Tame, J. R. H. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m412885200 · Cited by 180 · see citing works
Klemm, P., Hjerrild, L., Gjermansen, M., & Schembri, M. A. (2003) · Molecular Microbiology · doi:10.1046/j.1365-2958.2003.03833.x · Cited by 179 · see citing works
Self-citationSherlock, O., Schembri, M. A., Reisner, A., & Klemm, P. (2004) · Journal of Bacteriology · doi:10.1128/jb.186.23.8058-8065.2004 · Cited by 178 · see citing works
Self-citationAutotransporter structure reveals intra-barrel cleavage followed by conformational changes
Barnard, T. J., Dautin, N., Lukacik, P., et al. (2007) · Nature Structural & Molecular Biology · doi:10.1038/nsmb1322 · Cited by 165 · see citing works
Antigen 43 facilitates formation of multispecies biofilms
Kjærgaard, K., Schembri, M. A., Ramos, C., Molin, S., & Klemm, P. (2000) · Environmental Microbiology · doi:10.1046/j.1462-2920.2000.00152.x · Cited by 161 · see citing works
Self-citationCrystal Structure of a Full-Length Autotransporter
van den Berg, B. (2010) · Journal of Molecular Biology · doi:10.1016/j.jmb.2009.12.061 · Cited by 161 · see citing works
Crystal structure of the Helicobacter pylori vacuolating toxin p55 domain
Gangwer, K. A., Mushrush, D. J., Stauff, D. L., et al. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0707447104 · Cited by 161 · see citing works
Evolution of the β-barrel assembly machinery
Webb, C. T., Heinz, E., & Lithgow, T. J. (2012) · Trends in Microbiology · doi:10.1016/j.tim.2012.08.006 · Cited by 157 · see citing works
Henderson, I. R., Meehan, M., & Owen, P. (2006) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.1997.tb10317.x · Cited by 135 · see citing works
Uropathogenic Escherichia coli Mediated Urinary Tract Infection
Totsika, M., Moriel, D. G., Idris, A., Rogers, B. A., Wurpel, D. J., Phan, M., Paterson, D. L., & Schembri, M. A. (2012) · Current Drug Targets · doi:10.2174/138945012803530206 · Cited by 132 · see citing works
Self-citationDifferential Expression of the <i>Escherichia</i> <i>coli</i> Autoaggregation Factor Antigen 43
Schembri, M. A., Hjerrild, L., Gjermansen, M., & Klemm, P. (2003) · Journal of Bacteriology · doi:10.1128/jb.185.7.2236-2242.2003 · Cited by 126 · see citing works
Self-citationAutotransporter proteins, evolution and redefining protein secretion
Henderson, I. R., Cappello, R. E., & Nataro, J. P. (2000) · Trends in Microbiology · doi:10.1016/s0966-842x(00)01853-9 · Cited by 120 · see citing works
Statistical Analysis of Interface Similarity in Crystals of Homologous Proteins
Xu, Q., Canutescu, A. A., Wang, G., Shapovalov, M. V., Zoran Obradovic, A., & Dunbrack, R. L. (2008) · Journal of Molecular Biology · doi:10.1016/j.jmb.2008.06.002 · Cited by 102 · see citing works
Kjærgaard, K., Schembri, M. A., Hasman, H., & Klemm, P. (2000) · Journal of Bacteriology · doi:10.1128/jb.182.17.4789-4796.2000 · Cited by 101 · see citing works
Self-citationAutotransporter secretion: varying on a theme
Grijpstra, J., Arenas, J., Rutten, L., & Tommassen, J. (2013) · Research in Microbiology · doi:10.1016/j.resmic.2013.03.010 · Cited by 95 · see citing works
Kajava, A. V., & Steven, A. C. (2006) · Journal of Structural Biology · doi:10.1016/j.jsb.2006.01.015 · Cited by 95 · see citing works
Celik, N., Webb, C. T., Leyton, D. L., Holt, K. E., Heinz, E., Gorrell, R. J., Kwok, T., Naderer, T., Strugnell, R. A., Speed, T. P., et al. (2012) · PLoS ONE · doi:10.1371/journal.pone.0043245 · Cited by 86 · see citing works
Structures and functions of autotransporter proteins in microbial pathogens
Benz, I., & Schmidt, M. A. (2011) · International Journal of Medical Microbiology · doi:10.1016/j.ijmm.2011.03.003 · Cited by 86 · see citing works
Dehydration Converts DsbG Crystal Diffraction from Low to High Resolution
Heras, B., Edeling, M. A., Byriel, K. A., Jones, A., Raina, S., & Martin, J. L. (2003) · Structure · doi:10.1016/s0969-2126(03)00005-4 · Cited by 81 · see citing works
Self-citationA Novel Intein-Like Autoproteolytic Mechanism in Autotransporter Proteins
Tajima, N., Kawai, F., Park, S., et al. (2010) · Journal of Molecular Biology · doi:10.1016/j.jmb.2010.06.068 · Cited by 77 · see citing works
Meng, G., Spahich, N., Kenjale, R., Waksman, G., & St Geme, J. W. (2011) · The EMBO Journal · doi:10.1038/emboj.2011.279 · Cited by 76 · see citing works
Active-Site Gating Regulates Substrate Selectivity in a Chymotrypsin-Like Serine Protease
Johnson, T. A., Qiu, J., Plaut, A. G., & Holyoak, T. (2009) · Journal of Molecular Biology · doi:10.1016/j.jmb.2009.04.041 · Cited by 74 · see citing works
Autotransporters of Escherichia coli: a sequence-based characterization
Wells, T. J., Totsika, M., & Schembri, M. A. (2010) · Microbiology · doi:10.1099/mic.0.039024-0 · Cited by 63 · see citing works
Self-citationComplete fiber structures of complex trimeric autotransporter adhesins conserved in enterobacteria
Hartmann, M. D., Grin, I., Dunin-Horkawicz, S., et al. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1211872110 · Cited by 59 · see citing works
Acellular pertussis vaccines and the role of pertactin and fimbriae
POOLMAN, J. T., & Hallander, H. O. (2007) · Expert Review of Vaccines · doi:10.1586/14760584.6.1.47 · Cited by 57 · see citing works
Roche, A. J., McFadden, J. P., & Owen, P. (2001) · Microbiology · doi:10.1099/00221287-147-1-161 · Cited by 50 · see citing works
Crystal Structure of the Passenger Domain of the Escherichia coli Autotransporter EspP
Khan, S., Mian, H. S., Sandercock, L. E., et al. (2011) · Journal of Molecular Biology · doi:10.1016/j.jmb.2011.09.028 · Cited by 47 · see citing works
Zhai, Y., Zhang, K., Huo, Y., et al. (2011) · Biochemical Journal · doi:10.1042/bj20101548 · Cited by 46 · see citing works
Recombinogenic engineering of conjugative plasmids with fluorescent marker cassettes
Reisner, A., Molin, S., & Zechner, E. L. (2002) · FEMS Microbiology Ecology · doi:10.1111/j.1574-6941.2002.tb01015.x · Cited by 31 · see citing works
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Hobley, L., Harkins, C., MacPhee, C. E., & Stanley-Wall, N. R. (2015) · FEMS Microbiology Reviews · doi:10.1093/femsre/fuv015 · Cited by 581 · see citing works
Trunk, T., Bacterial Cell Surface Group, S. F. G. A. E. B., S. Khalil, H., & C. Leo, J. (2018) · AIMS Microbiology · doi:10.3934/microbiol.2018.1.140 · Cited by 444 · see citing works
“The model is based on the Ag43 crystal structure (PDB: 4KH3) [77].”
BackgroundMethodInfluentialChemotaxis towards autoinducer 2 mediates autoaggregation in Escherichia coli
Laganenka, L., Colin, R., & Sourjik, V. (2016) · Nature Communications · doi:10.1038/ncomms12984 · Cited by 218 · see citing works
Arenas, J., Cano, S., Nijland, R., van Dongen, V., Rutten, L., van der Ende, A., & Tommassen, J. (2015) · Environmental Microbiology · doi:10.1111/1462-2920.12581 · Cited by 43 · see citing works
Self-association motifs in the enteroaggregative Escherichia coli heat-resistant agglutinin 1
Glaubman, J., Hofmann, J., Bonney, M. E., Park, S., Thomas, J. M., Kokona, B., Ramos Falcón, L. I., Chung, Y. K., Fairman, R., & Okeke, I. N. (2016) · Microbiology · doi:10.1099/mic.0.000303 · Cited by 13 · see citing works
- Authors
- Heras, B., Totsika, M., Peters, K. M., Paxman, J. J., Gee, C. L., Jarrott, R. J., Perugini, M. A., Whitten, A. E., & Schembri, M. A.
- Affiliations
- La Trobe University (AU)
- The University of Queensland (AU)
- Australian Synchrotron (AU)
- ORCID
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2013-12-13
- Volume
- 111
- Issue
- 1
- Pages
- 457-462
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC3890832
- OpenAlex
- W2046216701
- Cited by
- 141
- References
- 49
- Citations by year
- 2026: 7 · 2025: 10 · 2024: 11 · 2023: 13 · 2022: 9 · 2021: 10 · 2020: 11 · 2019: 20 · 2018: 11 · 2017: 5 · 2016: 13 · 2015: 17 · 2014: 4
- FWCI
- 5.08
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- other-oa
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- QUT ePrints (Queensland University of Technology) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Escherichia coli research studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- antigen 43, autotransporter proteins, bacterial aggregation, self-association, uropathogenic Escherichia coli, biofilm formation, outer membrane proteins, structure-function relationship
- MeSH terms
- Antigens, Bacterial, Biological Transport, Cloning, Molecular, Hydrogen Bonding, Mutation, Protein Binding, Urinary Tract Infections, X-Ray Diffraction, Protein Structure, Secondary, Protein Structure, Tertiary, Crystallography, X-Ray, Biofilms, Adhesins, Bacterial, Adhesins, Escherichia coli, Uropathogenic Escherichia coli
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