Structural Basis of SspB-tail Recognition by the Zinc Binding Domain of ClpX

Eun Young Park; Byung Gil Lee; Seung Beom Hong; Hyung Wook Kim; Hyesung Jeon; Hyun Kyu Song

doi:10.1016/j.jmb.2007.01.003

Structural Basis of SspB-tail Recognition by the Zinc Binding Domain of ClpX

Eun Young Park, Byung Gil Lee, Seung Beom Hong, Hyung Wook Kim, Hyesung Jeon, Hyun Kyu Song

Research output: Contribution to journal › Article › peer-review

39 Citations (Scopus)

Abstract

The degradation of ssrA(AANDENYALAA)-tagged proteins in the bacterial cytosol is carried out by the ClpXP protease and is markedly stimulated by the SspB adaptor protein. It has previously been reported that the amino-terminal zinc-binding domain of ClpX (ZBD) is involved in complex formation with the SspB-tail (XB: ClpX-binding motif). In an effort to better understand the recognition of SspB by ClpX and the mechanism of delivery of ssrA-tagged substrates to ClpXP, we have determined the structures of ZBD alone at 1.5, 2.0, and 2.5 Å resolution in each different crystal form and also in complex with XB peptide at 1.6 Å resolution. The XB peptide forms an antiparallel β-sheet with two β-strands of ZBD, and the structure shows a 1:1 stoichiometric complex between ZBD and XB, suggesting that there are two independent SspB-tail-binding sites in ZBD. The high-resolution ZBD:XB complex structure, in combination with biochemical analyses, can account for key determinants in the recognition of the SspB-tail by ClpX and sheds light on the mechanism of delivery of target proteins to the prokaryotic degradation machine.

Original language	English
Pages (from-to)	514-526
Number of pages	13
Journal	Journal of Molecular Biology
Volume	367
Issue number	2
DOIs	https://doi.org/10.1016/j.jmb.2007.01.003
Publication status	Published - 2007 Mar 23

Bibliographical note

Funding Information:
We thank the staff at 4A beamline, Pohang Accelerator Laboratory, Republic of Korea and NW12 beamline, Photon Factory, Japan for help with data collection. We also thank Drs P. Zwickl (Max-Planck-Institute for Biochemistry, Germany) and T. Tamura (National Institute of Advanced Industrial Science and Technology, Japan) for cells containing His-GFP-ssrA and Professor M. J. Eck for his generous support during the initial stage of this project. This work was supported by the Korea Research Foundation Grant (KRF-2006-312-C00249) funded by the Korean Government (MOEHRD) and by the Korea Science and Engineering Foundation Grant (KOSEF, R01-2004-000-10773-0) funded by the Korea government (MOST). E.Y.P. was supported by a Seoul Science Fellowship.

Keywords

ATP-dependent protease
ClpXP
crystal
delivery complex
ssrA tag

ASJC Scopus subject areas

Biophysics
Structural Biology
Molecular Biology

Access to Document

10.1016/j.jmb.2007.01.003

Cite this

@article{7ae8b7a7add542d19c80d0e1394a04d9,

title = "Structural Basis of SspB-tail Recognition by the Zinc Binding Domain of ClpX",

abstract = "The degradation of ssrA(AANDENYALAA)-tagged proteins in the bacterial cytosol is carried out by the ClpXP protease and is markedly stimulated by the SspB adaptor protein. It has previously been reported that the amino-terminal zinc-binding domain of ClpX (ZBD) is involved in complex formation with the SspB-tail (XB: ClpX-binding motif). In an effort to better understand the recognition of SspB by ClpX and the mechanism of delivery of ssrA-tagged substrates to ClpXP, we have determined the structures of ZBD alone at 1.5, 2.0, and 2.5 {\AA} resolution in each different crystal form and also in complex with XB peptide at 1.6 {\AA} resolution. The XB peptide forms an antiparallel β-sheet with two β-strands of ZBD, and the structure shows a 1:1 stoichiometric complex between ZBD and XB, suggesting that there are two independent SspB-tail-binding sites in ZBD. The high-resolution ZBD:XB complex structure, in combination with biochemical analyses, can account for key determinants in the recognition of the SspB-tail by ClpX and sheds light on the mechanism of delivery of target proteins to the prokaryotic degradation machine.",

keywords = "ATP-dependent protease, ClpXP, crystal, delivery complex, ssrA tag",

author = "Park, {Eun Young} and Lee, {Byung Gil} and Hong, {Seung Beom} and Kim, {Hyung Wook} and Hyesung Jeon and Song, {Hyun Kyu}",

note = "Funding Information: We thank the staff at 4A beamline, Pohang Accelerator Laboratory, Republic of Korea and NW12 beamline, Photon Factory, Japan for help with data collection. We also thank Drs P. Zwickl (Max-Planck-Institute for Biochemistry, Germany) and T. Tamura (National Institute of Advanced Industrial Science and Technology, Japan) for cells containing His-GFP-ssrA and Professor M. J. Eck for his generous support during the initial stage of this project. This work was supported by the Korea Research Foundation Grant (KRF-2006-312-C00249) funded by the Korean Government (MOEHRD) and by the Korea Science and Engineering Foundation Grant (KOSEF, R01-2004-000-10773-0) funded by the Korea government (MOST). E.Y.P. was supported by a Seoul Science Fellowship.",

year = "2007",

month = mar,

day = "23",

doi = "10.1016/j.jmb.2007.01.003",

language = "English",

volume = "367",

pages = "514--526",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - Structural Basis of SspB-tail Recognition by the Zinc Binding Domain of ClpX

AU - Park, Eun Young

AU - Lee, Byung Gil

AU - Hong, Seung Beom

AU - Kim, Hyung Wook

AU - Jeon, Hyesung

AU - Song, Hyun Kyu

N1 - Funding Information: We thank the staff at 4A beamline, Pohang Accelerator Laboratory, Republic of Korea and NW12 beamline, Photon Factory, Japan for help with data collection. We also thank Drs P. Zwickl (Max-Planck-Institute for Biochemistry, Germany) and T. Tamura (National Institute of Advanced Industrial Science and Technology, Japan) for cells containing His-GFP-ssrA and Professor M. J. Eck for his generous support during the initial stage of this project. This work was supported by the Korea Research Foundation Grant (KRF-2006-312-C00249) funded by the Korean Government (MOEHRD) and by the Korea Science and Engineering Foundation Grant (KOSEF, R01-2004-000-10773-0) funded by the Korea government (MOST). E.Y.P. was supported by a Seoul Science Fellowship.

PY - 2007/3/23

Y1 - 2007/3/23

N2 - The degradation of ssrA(AANDENYALAA)-tagged proteins in the bacterial cytosol is carried out by the ClpXP protease and is markedly stimulated by the SspB adaptor protein. It has previously been reported that the amino-terminal zinc-binding domain of ClpX (ZBD) is involved in complex formation with the SspB-tail (XB: ClpX-binding motif). In an effort to better understand the recognition of SspB by ClpX and the mechanism of delivery of ssrA-tagged substrates to ClpXP, we have determined the structures of ZBD alone at 1.5, 2.0, and 2.5 Å resolution in each different crystal form and also in complex with XB peptide at 1.6 Å resolution. The XB peptide forms an antiparallel β-sheet with two β-strands of ZBD, and the structure shows a 1:1 stoichiometric complex between ZBD and XB, suggesting that there are two independent SspB-tail-binding sites in ZBD. The high-resolution ZBD:XB complex structure, in combination with biochemical analyses, can account for key determinants in the recognition of the SspB-tail by ClpX and sheds light on the mechanism of delivery of target proteins to the prokaryotic degradation machine.

AB - The degradation of ssrA(AANDENYALAA)-tagged proteins in the bacterial cytosol is carried out by the ClpXP protease and is markedly stimulated by the SspB adaptor protein. It has previously been reported that the amino-terminal zinc-binding domain of ClpX (ZBD) is involved in complex formation with the SspB-tail (XB: ClpX-binding motif). In an effort to better understand the recognition of SspB by ClpX and the mechanism of delivery of ssrA-tagged substrates to ClpXP, we have determined the structures of ZBD alone at 1.5, 2.0, and 2.5 Å resolution in each different crystal form and also in complex with XB peptide at 1.6 Å resolution. The XB peptide forms an antiparallel β-sheet with two β-strands of ZBD, and the structure shows a 1:1 stoichiometric complex between ZBD and XB, suggesting that there are two independent SspB-tail-binding sites in ZBD. The high-resolution ZBD:XB complex structure, in combination with biochemical analyses, can account for key determinants in the recognition of the SspB-tail by ClpX and sheds light on the mechanism of delivery of target proteins to the prokaryotic degradation machine.

KW - ATP-dependent protease

KW - ClpXP

KW - crystal

KW - delivery complex

KW - ssrA tag

UR - http://www.scopus.com/inward/record.url?scp=33847108004&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2007.01.003

DO - 10.1016/j.jmb.2007.01.003

M3 - Article

C2 - 17258768

AN - SCOPUS:33847108004

SN - 0022-2836

VL - 367

SP - 514

EP - 526

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 2

ER -

Structural Basis of SspB-tail Recognition by the Zinc Binding Domain of ClpX

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this