Chinese Journal of Magnetic Resonance >
Solution Structure of Bacillus subtilis Twin-Arginine Translocation TatAy Protein
Received date: 2015-02-09
Revised date: 2015-05-09
Online published: 2015-06-05
Supported by
Grant from the Ministry of Science and Technology of China (2010IM030700) and the National Natural Science Foundation of China (31070649).
The twin-arginine transport (Tat) systems in bacteria and plant chloroplasts translocate cargo proteins across cellular membranes in their folded states. The single-pass transmembrane protein TatA forms the protein translocation channel via self-oligomerization. Herein, we present the structure of Bacillus subtilis TatAy protein in dodecylphosphocholine micelles determined by solution NMR method. TatAy adopts an L-shaped conformation formed by a transmembrane helix (TMH) and an amphipathic helix (APH). Structural comparison of TatAy protein with the previously reported B. subtilis TatAd protein highlights essential residues at the hinge region for maintaining the L-shaped conformation, and suggests a few conserved structural features for the
TatA protein family. Possible roles for the conserved residues in the TatA channel formation are discussed.
HU Yun-fei1,2，HE Peng3，WU Yu-jie1,3，JIN Chang-wen1,2,3,4* . Solution Structure of Bacillus subtilis Twin-Arginine Translocation TatAy Protein[J]. Chinese Journal of Magnetic Resonance, 2015 , 32(2) : 291 -307 . DOI: 10.11938/cjmr20150212
[1] Palmer T, Berks B C. The twin-arginine translocation (Tat) protein export pathway[J]. Nat Rev Micro, 2012, 10(7): 483－496.
[2] Sargent F. The twin-arginine transport system: moving folded proteins across membranes[J]. Biochem Soc Trans, 2007, 35(Pt 5): 835－847.
[3] Weiner J H, Bilous P T, Shaw G M, et al. A novel and ubiquitous system for membrane targeting and secretion of cofactor-containing proteins[J]. Cell, 1998, 93(1): 93－101.
[4] Sargent F, Bogsch E G, Stanley N R, et al. Overlapping functions of components of a bacterial Sec-independent protein export pathway[J]. EMBO J, 1998, 17(13): 3 640－3 650.
[5] Bogsch E G, Sargent F, Stanley N R, et al. An essential component of a novel bacterial protein export system with homologues in plastids and mitochondria[J]. J Biol Chem, 1998, 273(29): 18 003－18 006.
[6] Sargent F, Stanley N R, Berks, B C, et al. Sec-independent protein translocation in Escherichia coli. A distinct and pivotal role for the TatB protein[J]. J Biol Chem, 1999, 274(51): 36 073－36 082.
[7] Jongbloed J D, Grieger U, Antelmann H, et al. Two minimal Tat translocases in Bacillus[J]. Mol Microbiol, 2004, 54(5): 1 319－1 325.
[8] Gohlke U, Pullan L, McDevitt C A, et al. The TatA component of the twin-arginine protein transport system forms channel complexes of variable diameter[J]. Proc Natl Acad Sci USA, 2005, 102(30): 10 482－10 486.
[9] Dabney-Smith C, Mori H, Cline K. Oligomers of Tha4 organize at the thylakoid Tat translocase during protein transport[J]. J Biol Chem, 2006, 281(9): 5 476－5 483.
[10] Leake M C, Greene N P, Godun R M, et al. Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging[J]. Proc Natl Acad Sci USA, 2008, 105(40): 15 376－15 381.
[11] de Leeuw E, Granjon T, Porcelli I, et al. Oligomeric properties and signal peptide binding by Escherichia coli Tat protein transport complexes[J]. J Mol Biol, 2002, 322(5): 1 135－1 146.
[12] Cline K, Mori H. Thylakoid deltapH-dependent precursor proteins bind to a cpTatC-Hcf106 complex before Tha4-dependent transport[J]. J Cell Biol, 2001, 154(4): 719－729.
[13] Alami M, Lüke I, Deitermann S, et al. Differential interactions between a twin-arginine signal peptide and its translocase in Escherichia coli[J]. Mol Cell, 2003, 12(4): 937－946.
[14] Yahr T L, Wickner W T. Functional reconstitution of bacterial Tat translocation in vitro[J]. EMBO J, 2001, 20(10): 2 472－2 479.
[15] Westermann M, Pop O I, Gerlach R, et al. The TatAd component of the Bacillus subtilis twin-arginine protein transport system forms homo-multimeric complexes in its cytosolic and membrane embedded localization[J]. Biochim Biophys Acta, 2006, 1 758(4): 443－451.
[16] Barnett J P, Eijlander R T, Kuipers O P, et al. A minimal Tat system from a gram-positive organism: a bifunctional TatA subunit participates in discrete TatAC and TatA complexes[J]. J Biol Chem, 2006, 283(5): 2 534－2 542.
[17] Pop O I, Westermann M, Volkmer-Engert R, et al. Sequence-specific binding of prePhoD to soluble TatAd indicates protein-mediated targeting of the Tat export in Bacillus subtilis[J]. J Biol Chem, 2003, 278(40): 38 428－38 436.
[18] Rollauer S E, Tarry M J, Graham J E, et al. Structure of the TatC core of the twin-arginine protein transport system[J]. Nature, 2012, 492(7428): 210－214.
[19] Ramasamy S, Abrol R, Suloway C J, et al. The glove-like structure of the conserved membrane protein TatC provides insight into signal sequence recognition in twin-arginine translocation[J]. Structure, 2013, 21(5): 777－788.
[20] Rodriguez F, Rouse S L, Tait C E, et al. Structural model for the protein-translocating element of the twin-arginine transport system[J]. Proc Natl Acad Sci USA, 2013, 110(12): E1 092－E1 101.
[21] Zhang Y, Hu Y, Li H, et al. Structural basis for TatA oligomerization: an NMR study of Escherichia coli TatA dimeric structure[J]. PLoS One, 2014, 9(8): e103157
[22] Zhang Y, Wang L, Hu Y, et al. Solution structure of the TatB component of the twin-arginine translocation system[J]. Biochim Biophys Acta, 2014, 1838(7): 1 881－1 888.
[23] Hu Y, Zhao E, Li H, et al. Solution NMR structure of the TatA component of the twin-arginine protein transport system from gram-positive bacterium Bacillus subtilis[J]. J Am Chem Soc, 2010, 132(45): 15 942－15 944.
[24] Walther T H, Grage S L, Roth N, et al. Membrane alignment of the pore-forming component TatAd of the twin-arginine translocase from Bacillus subtilis resolved by solid-state NMR spectroscopy[J]. J Am Chem Soc, 2010, 132(45): 15 945－15 956.
[25] Walther T H, Gottselig C, Grage S L, et al. Folding and self-assembly of the TatA translocation pore based on a charge zipper mechanism[J]. Cell, 2013, 152(1-2): 316－326.
[26] Pop O, Martin U, Abel C, et al. The twin-arginine signal peptide of PhoD and the TatAd/Cd proteins of Bacillus subtilis form an autonomous Tat translocation system[J]. J Biol Chem, 2002, 277(5): 3 268－3 273.
[27] Sattler M, Schleucher J, Griesinger C. Heteronuclear multidimensional NMR experiments for the structure determination of proteins in solution employing pulse field gradients[J]. Prog Nucl Magn Reson Spect, 1999, 34: 93－158.
[28] Delaglio F, Grzesiek S, Vuister G W, et al. NMRPipe: a multidimensional spectral processing system based on UNIX pipes[J]. J Biomol NMR, 1995, 6(3): 277－293.
[29] Johnson B A. Using NMRView to visualize and analyze the NMR spectra of macromolecules[J]. Methods Mol Biol, 2004, 278: 313－352.
[30] Lorieau J, Yao L, Bax A. Liquid crystalline phase of G-tetrad DNA for NMR study of detergent-solubilized proteins[J]. J Am Chem Soc, 2008, 130(24): 7 536－7 537.
[31] Ottiger M, Delaglio F, Bax A. Measurement of J and dipolar couplings from simplified two-dimensional NMR spectra[J]. J. Magn. Reson. 1998, 131(2): 373－378.
[32] Zweckstetter M. NMR: prediction of molecular alignment from structure using the PALES software[J]. Nat Protoc, 2008, 3(4): 679－690.
[33] Dosset P, Hus J C, Marion D, et al. A novel interactive tool for rigid-body modeling of multi-domain macromolecules using residual dipolar couplings[J]. J Biomol NMR, 2001, 20(3): 223－231.
[34] Cornilescu G, Delaglio F, Bax, A. Protein backbone angle restraints from searching a database for chemical shift and sequence homology[J]. J Biomol NMR, 1999, 13(3): 289－302.
[35] Schwieters C D, Kuszewski J J, Tjandra N, et al. The Xplor-NIH NMR molecular structure determination package[J]. J Magn Reson, 2003, 160(1): 65－73.
[36] Farrow N A, Muhandiram R, Singer A U, et al. Backbone dynamics of a free and phosphopeptide-complexed Src homology 2 domain studied by 15N NMR relaxation[J]. Biochemistry, 1994, 33(19): 5 984－6 003.
[37] Barrett C M, Mathers J E, Robinson C. Identification of key regions within the Escherichia coli TatAB subunits[J]. FEBS Lett, 2003, 537(1-3): 42－46.
[38] Hicks M G, de Leeuw E, Porcelli I, et al. The Escherichia coli twin-arginine translocase: conserved residues of TatA and TatB family components involved in protein transport[J]. FEBS Lett, 2003, 539(1-3): 61－67.
[39] White G F, Schermann S M, Bradley J, et al. Subunit organization in the TatA complex of the twin arginine protein translocase: a site-directed EPR spin labeling study[J]. J Biol Chem, 2010, 285(4): 2 294－2 301.
[40] Greene N P, Porcelli I, Buchanan G, et al. Cysteine scanning mutagenesis and disulfide mapping studies of the TatA component of the bacterial twin arginine translocase[J]. J Biol Chem, 2007, 282(33): 23 937－23 945.
[41] Barrett C M L, Mangels D, Robinson C. Mutations in subunits of the Escherichia coli twin-arginine translocase block function via differing effects on translocation activity or tat complex structure[J]. J Mol Biol, 2005, 347(2): 453－463.
[42] Hicks M G, Lee P A, Georgiou G, et al. Positive selection for loss-of-function tat mutations identifies critical residues required for TatA activity[J.] J Bacteriol, 2005, 187(8): 2 920－2 925.
[43] Gouffi K, Gérard F, Santini C L, et al. Dual topology of the Escherichia coli TatA protein[J]. J Biol Chem, 2004, 279(12): 11 608－11 615.
[44] Chan C S, Zlomislic M R, Tieleman D P, et al. The TatA subunit of Escherichia coli twin-arginine translocase has an N-in topology[J]. Biochemistry, 2007, 46(25): 7 396－7 404.
[45] Behrendt J, Standar K, Lindenstrauss U, et al. Topological studies on the twin-arginine translocase component TatC[J]]. FEMS Microbiol Lett, 2004, 234(2): 303－308.
[46] Gouffi K, Santini C L, Wu L F. Topology determination and functional analysis of the Escherichia coli TatC protein[J]. FEBS Lett, 2002, 525(1-3): 65－70.
[47] Kneller J M, Lu M, Bracken C. An effective method for the discrimination of motional anisotropy and chemical exchange[J]. J Am Chem Soc, 2002, 124(9): 1 852－1 853.
[48] Lange C, Müller S D, Walther T H, et al. Structure analysis of the protein translocating channel TatA in membranes using a multi-construct approach[J]. Biochim Biophys Acta, 2007, 1 768(10): 2 627－2 634.
[49] Porcelli I, de Leeuw E, Wallis R, et al. Characterization and membrane assembly of the TatA component of the Escherichia coli twin-arginine protein transport system[J]. Biochemistry, 2002, 41(46): 13 690－13 697.
[50] Mikhaleva N I, Santini C L, Giordano G, et al. Requirement for phospholipids of the translocation of the trimethylamine N-oxide reductase through the Tat pathway in Escherichia coli[J]. FEBS Lett, 1999, 463(3): 331－335.
/
| 〈 |
|
〉 |