Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii

Vered Irihimovitch; Gabriela Ring; Tsiona Elkayam; Zvia Konrad; Jerry Eichler

doi:10.1007/s00792-002-0297-0

Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii

Vered Irihimovitch, Gabriela Ring, Tsiona Elkayam, Zvia Konrad, Jerry Eichler

Department of Life Sciences

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

19 Scopus citations

Abstract

By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE.

Original language	English
Pages (from-to)	71-77
Number of pages	7
Journal	Extremophiles
Volume	7
Issue number	1
DOIs	https://doi.org/10.1007/s00792-002-0297-0
State	Published - 1 Dec 2003

Keywords

Archaea
Cellulose-binding domain
Haloferax volcanii
Halophiles
SecE
SecY

ASJC Scopus subject areas

Microbiology
Molecular Medicine

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title = "Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii",

abstract = "By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE.",

keywords = "Archaea, Cellulose-binding domain, Haloferax volcanii, Halophiles, SecE, SecY",

author = "Vered Irihimovitch and Gabriela Ring and Tsiona Elkayam and Zvia Konrad and Jerry Eichler",

note = "Funding Information: Acknowledgments The authors wish to thank Dr. Moshe Mevar-ech for strains, antibodies, and advice and Dr. Arie Admon for antibodies. This work was supported by the Israel Ministry of Absorption and the Israel Science Foundation (grant no. 291/99). J.E. is the incumbent of the Murray Shusterman Career Development Chair in Microbiology.",

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doi = "10.1007/s00792-002-0297-0",

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T1 - Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii

AU - Irihimovitch, Vered

AU - Ring, Gabriela

AU - Elkayam, Tsiona

AU - Konrad, Zvia

AU - Eichler, Jerry

N1 - Funding Information: Acknowledgments The authors wish to thank Dr. Moshe Mevar-ech for strains, antibodies, and advice and Dr. Arie Admon for antibodies. This work was supported by the Israel Ministry of Absorption and the Israel Science Foundation (grant no. 291/99). J.E. is the incumbent of the Murray Shusterman Career Development Chair in Microbiology.

PY - 2003/12/1

Y1 - 2003/12/1

N2 - By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE.

AB - By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE.

KW - Archaea

KW - Cellulose-binding domain

KW - Haloferax volcanii

KW - Halophiles

KW - SecE

KW - SecY

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DO - 10.1007/s00792-002-0297-0

M3 - Article

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SN - 1431-0651

VL - 7

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EP - 77

JO - Extremophiles

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IS - 1

ER -

Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii

Abstract

Keywords

ASJC Scopus subject areas

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