EsxB, a secreted protein from Bacillus anthracis forms two distinct helical bundles

Fan, Yao; Tan, Kemin; Chhor, Gekleng; Butler, Emily K.; Jedrzejczak, Robert P.; Missiakas, Dominique; Joachimiak, Andrzej

doi:10.1002/pro.2715

EsxB, a secreted protein from Bacillus anthracis forms two distinct helical bundles

Journal Article · Fri Jul 03 00:00:00 EDT 2015 · Protein Science

DOI:https://doi.org/10.1002/pro.2715· OSTI ID:1212699

Fan, Yao ^[1]; Tan, Kemin ^[2]; Chhor, Gekleng ^[1]; Butler, Emily K. ^[2]; Jedrzejczak, Robert P. ^[1]; Missiakas, Dominique ^[2]; Joachimiak, Andrzej ^[2]

Argonne National Lab. (ANL), Argonne, IL (United States)
Argonne National Lab. (ANL), Argonne, IL (United States); Univ. of Chicago, Chicago, IL (United States)

The EsxB protein from Bacillus anthracis belongs to the WXG100 family, a group of proteins secreted by a specialized secretion system. We have determined the crystal structures of recombinant EsxB and discovered that the small protein (~10 kDa), comprised of a helix-loop-helix (HLH) hairpin, is capable of associating into two different helical bundles. The two basic quaternary assemblies of EsxB are an antiparallel (AP) dimer and a rarely observed bisecting U (BU) dimer. This structural duality of EsxB is believed to originate from the heptad repeat sequence diversity of the first helix of its HLH hairpin, which allows for two alternative helix packing. The flexibility of EsxB and the ability to form alternative helical bundles underscore the possibility that this protein can serve as an adaptor in secretion and can form hetero-oligomeric helix bundle(s) with other secreted members of the WXG100 family, such as EsxW. The highly conserved WXG motif is located within the loop of the HLH hairpin and is mostly buried within the helix bundle suggesting that its role is mainly structural. The exact functions of the motif, including a proposed role as a secretion signal, remain unknown.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1212699

Alternate ID(s):: OSTI ID: 1391889

Journal Information:: Protein Science, Journal Name: Protein Science Journal Issue: 9 Vol. 24; ISSN 0961-8368

Publisher:: The Protein SocietyCopyright Statement

Country of Publication:: United States

Language:: English

References (33)

A structural mechanism for dimeric to tetrameric oligomer conversion in Halomonas sp. nucleoside diphosphate kinase: Oligomer Conversion Mechanism of HaNDK Arai, Shigeki; Yonezawa, Yasushi; Okazaki, Nobuo Protein Science, Vol. 21, Issue 4 https://doi.org/10.1002/pro.2032	journal	March 2012
Atomic Structures of the Human Immunophilin FKBP-12 Complexes with FK506 and Rapamycin Van Duyne, Gregory D.; Standaert, Robert F.; Karplus, P. Andrew Journal of Molecular Biology, Vol. 229, Issue 1 https://doi.org/10.1006/jmbi.1993.1012	journal	January 1993
The Polymerase Incomplete Primer Extension (PIPE) Method Applied to High-Throughput Cloning and Site-Directed Mutagenesis Klock, Heath E.; Lesley, Scott A. Methods in Molecular Biology https://doi.org/10.1007/978-1-59745-196-3_6	book	January 2009
New LIC vectors for production of proteins from genes containing rare codons Eschenfeldt, William H.; Makowska-Grzyska, Magdalena; Stols, Lucy Journal of Structural and Functional Genomics, Vol. 14, Issue 4 https://doi.org/10.1007/s10969-013-9163-9	journal	September 2013
The ESAT-6/WXG100 superfamily – and a new Gram-positive secretion system? Pallen, Mark J. Trends in Microbiology, Vol. 10, Issue 5 https://doi.org/10.1016/S0966-842X(02)02345-4	journal	May 2002
Protein plasticity to the extreme: changing the topology of a 4-α-helical bundle with a single amino acid substitution Glykos, Nicholas M.; Cesareni, Gianni; Kokkinidis, Michael Structure, Vol. 7, Issue 6 https://doi.org/10.1016/S0969-2126(99)80081-1	journal	June 1999
The effect of the hexahistidine-tag in the oligomerization of HSC70 constructs Amor-Mahjoub, Mouna; Suppini, Jean-Philippe; Gomez-Vrielyunck, Nathalie Journal of Chromatography B, Vol. 844, Issue 2 https://doi.org/10.1016/j.jchromb.2006.07.031	journal	December 2006
Inference of Macromolecular Assemblies from Crystalline State Krissinel, Evgeny; Henrick, Kim Journal of Molecular Biology, Vol. 372, Issue 3 https://doi.org/10.1016/j.jmb.2007.05.022	journal	September 2007
Structure of Staphylococcus aureus EsxA Suggests a Contribution to Virulence by Action as a Transport Chaperone and/or Adaptor Protein Sundaramoorthy, Ramasubramanian; Fyfe, Paul K.; Hunter, William N. Journal of Molecular Biology, Vol. 383, Issue 3 https://doi.org/10.1016/j.jmb.2008.08.047	journal	November 2008
Outer membrane translocation: numerical protein secretion nomenclature in question in mycobacteria Desvaux, Mickaël; Hébraud, Michel; Talon, Régine Trends in Microbiology, Vol. 17, Issue 8 https://doi.org/10.1016/j.tim.2009.05.008	journal	August 2009
Solution Structure of α ₂ D, a Nativelike de Novo Designed Protein Hill, R. Blake; DeGrado, William F. Journal of the American Chemical Society, Vol. 120, Issue 6 https://doi.org/10.1021/ja9733649	journal	February 1998
Automation of protein purification for structural genomics Kim, Youngchang; Dementieva, Irina; Zhou, Min Journal of Structural and Functional Genomics, Vol. 5, Issue 1/2 https://doi.org/10.1023/B:JSFG.0000029206.07778.fc	journal	March 2004
Type VII secretion — mycobacteria show the way Abdallah, Abdallah M.; Gey van Pittius, Nicolaas C.; DiGiuseppe Champion, Patricia A. Nature Reviews Microbiology, Vol. 5, Issue 11 https://doi.org/10.1038/nrmicro1773	journal	November 2007
A phenylalanine zipper mediates APS dimerization Dhe-Paganon, Sirano; Werner, Eric D.; Nishi, Masahiro Nature Structural & Molecular Biology, Vol. 11, Issue 10 https://doi.org/10.1038/nsmb829	journal	September 2004
Structure and function of the complex formed by the tuberculosis virulence factors CFP-10 and ESAT-6 Renshaw, Philip S.; Lightbody, Kirsty L.; Veverka, Vaclav The EMBO Journal, Vol. 24, Issue 14 https://doi.org/10.1038/sj.emboj.7600732	journal	June 2005
Symmetry and frustration in protein energy landscapes: A near degeneracy resolves the Rop dimer-folding mystery Levy, Y.; Cho, S. S.; Shen, T. Proceedings of the National Academy of Sciences, Vol. 102, Issue 7 https://doi.org/10.1073/pnas.0409572102	journal	February 2005
Acute infection and macrophage subversion by Mycobacterium tuberculosis require a specialized secretion system Stanley, S. A.; Raghavan, S.; Hwang, W. W. Proceedings of the National Academy of Sciences, Vol. 100, Issue 22 https://doi.org/10.1073/pnas.2235593100	journal	October 2003
The Crystal and Solution Structures of Glyceraldehyde-3-phosphate Dehydrogenase Reveal Different Quaternary Structures Ferreira-da-Silva, Frederico; Pereira, Pedro J. B.; Gales, Luís Journal of Biological Chemistry, Vol. 281, Issue 44 https://doi.org/10.1074/jbc.M605267200	journal	September 2006
CASTp: Computed Atlas of Surface Topography of proteins Binkowski, T. A. Nucleic Acids Research, Vol. 31, Issue 13 https://doi.org/10.1093/nar/gkg512	journal	July 2003
Protein secretion and surface display in Gram-positive bacteria Schneewind, Olaf; Missiakas, Dominique M. Philosophical Transactions of the Royal Society B: Biological Sciences, Vol. 367, Issue 1592 https://doi.org/10.1098/rstb.2011.0210	journal	April 2012
Substructure solution with SHELXD Schneider, Thomas R.; Sheldrick, George M. Acta Crystallographica Section D Biological Crystallography, Vol. 58, Issue 10 https://doi.org/10.1107/S0907444902011678	journal	September 2002
Coot model-building tools for molecular graphics Emsley, Paul; Cowtan, Kevin Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132 https://doi.org/10.1107/S0907444904019158	journal	November 2004
Towards complete validated models in the next generation of ARP / wARP Cohen, Serge X.; Morris, Richard J.; Fernandez, Francisco J. Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12 https://doi.org/10.1107/S0907444904027556	journal	November 2004
HKL -3000: the integration of data reduction and structure solution – from diffraction images to an initial model in minutes Minor, Wladek; Cymborowski, Marcin; Otwinowski, Zbyszek Acta Crystallographica Section D Biological Crystallography, Vol. 62, Issue 8 https://doi.org/10.1107/S0907444906019949	journal	July 2006
MolProbity : all-atom structure validation for macromolecular crystallography Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1 https://doi.org/10.1107/S0907444909042073	journal	December 2009
Molecular replacement with MOLREP Vagin, Alexei; Teplyakov, Alexei Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1 https://doi.org/10.1107/S0907444909042589	journal	December 2009
The CCP4 suite programs for protein crystallography No authors listed Acta Crystallographica Section D Biological Crystallography, Vol. 50, Issue 5, p. 760-763 https://doi.org/10.1107/S0907444994003112	journal	September 1994
Refinement of Macromolecular Structures by the Maximum-Likelihood Method Murshudov, G. N.; Vagin, A. A.; Dodson, E. J. Acta Crystallographica Section D Biological Crystallography, Vol. 53, Issue 3 https://doi.org/10.1107/S0907444996012255	journal	May 1997
The Structural Biology Center 19ID undulator beamline: facility specifications and protein crystallographic results Rosenbaum, Gerd; Alkire, Randy W.; Evans, Gwyndaf Journal of Synchrotron Radiation, Vol. 13, Issue 1 https://doi.org/10.1107/S0909049505036721	journal	December 2005
ESAT-6-Like Protein Secretion in Bacillus anthracis Garufi, Gabriella; Butler, Emily; Missiakas, Dominique Journal of Bacteriology, Vol. 190, Issue 21 https://doi.org/10.1128/JB.00458-08	journal	August 2008
Dimer-Tetramer Transition between Solution and Crystalline States of Streptavidin and Avidin Mutants Pazy, Yael; Eisenberg-Domovich, Yael; Laitinen, Olli H. Journal of Bacteriology, Vol. 185, Issue 14 https://doi.org/10.1128/JB.185.14.4050-4056.2003	journal	July 2003
WXG100 Protein Superfamily Consists of Three Subfamilies and Exhibits an α-Helical C-Terminal Conserved Residue Pattern Poulsen, Christian; Panjikar, Santosh; Holton, Simon J. PLoS ONE, Vol. 9, Issue 2 https://doi.org/10.1371/journal.pone.0089313	journal	February 2014
Hexahistidine (His6)-tag dependent protein dimerization: a cautionary tale. Wu, J.; Filutowicz, M. Acta Biochimica Polonica, Vol. 46, Issue 3 https://doi.org/10.18388/abp.1999_4131	journal	September 1999

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59 BASIC BIOLOGICAL SCIENCES
ESAT-6 like secretion system
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helix bundle
tetramer
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