Publication

Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E

Driessen, A. J. M., Brundage, L., Hendrick, J. P., Schiebel, E. & Wickner, W., 1991, Vectorial Pansport of Proteins into and across Membranes. Tartakoff, A. M. (ed.). Vol. 34. p. 147-165 19 p. (Methods in cell biology).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

APA

Driessen, A. J. M., Brundage, L., Hendrick, J. P., Schiebel, E., & Wickner, W. (1991). Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E. In A. M. Tartakoff (Ed.), Vectorial Pansport of Proteins into and across Membranes (Vol. 34, pp. 147-165). (Methods in cell biology). https://doi.org/10.1016/S0091-679X(08)61679-9

Author

Driessen, A. J. M. ; Brundage, L. ; Hendrick, Joseph P. ; Schiebel, E. ; Wickner, William. / Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E. Vectorial Pansport of Proteins into and across Membranes. editor / Alan M. Tartakoff. Vol. 34 1991. pp. 147-165 (Methods in cell biology).

Harvard

Driessen, AJM, Brundage, L, Hendrick, JP, Schiebel, E & Wickner, W 1991, Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E. in AM Tartakoff (ed.), Vectorial Pansport of Proteins into and across Membranes. vol. 34, Methods in cell biology, pp. 147-165. https://doi.org/10.1016/S0091-679X(08)61679-9

Standard

Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E. / Driessen, A. J. M.; Brundage, L.; Hendrick, Joseph P.; Schiebel, E.; Wickner, William.

Vectorial Pansport of Proteins into and across Membranes. ed. / Alan M. Tartakoff. Vol. 34 1991. p. 147-165 (Methods in cell biology).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Vancouver

Driessen AJM, Brundage L, Hendrick JP, Schiebel E, Wickner W. Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E. In Tartakoff AM, editor, Vectorial Pansport of Proteins into and across Membranes. Vol. 34. 1991. p. 147-165. (Methods in cell biology). https://doi.org/10.1016/S0091-679X(08)61679-9


BibTeX

@inbook{0eae07c4db8d4d13ab155cb58e6fc6f3,
title = "Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E",
abstract = "This chapter describes the procedures to solubilize and purify a functional secY/E protein that, upon reconstitution, supports an authentic translocation reaction of precursor proteins. The secY/E protein is purified from octylglucoside-extracted membranes by the ability of the reconstituted enzyme to stimulate the ATP-hydrolyzing activity of the purified secA protein in the presence of proOmpA. The chapter describes the reconstitution procedure and the various assays required to determine the activity of the reconstituted secY/E protein. Protocols used for the isolation and solubilization of E. coli inner membranes and the purification of the secY/E protein are described. The purified secY/E protein contains three major polypeptide species. These are identified by immunoblots with antisera to secY and secE and by N-terminal sequence analysis. The largest polypeptide of the purified protein reacts with antibodies to the secY N-terminus and migrates on SDS-PAGE with an apparent molecular mass of 29 kDa. The use of glycerol and phospholipids with octylglucoside has a dramatic effect on the solubility of the secY protein.",
keywords = "MALTOSE-BINDING PROTEIN, PRECURSOR PROTEINS, PLASMA-MEMBRANE, TRIGGER FACTOR, SECA PROTEIN, PRO-OMPA, FUNCTIONAL RECONSTITUTION, CYTOPLASMIC MEMBRANE, TRANSPORT-SYSTEM, GENE-PRODUCT",
author = "Driessen, {A. J. M.} and L. Brundage and Hendrick, {Joseph P.} and E. Schiebel and William Wickner",
year = "1991",
doi = "10.1016/S0091-679X(08)61679-9",
language = "English",
isbn = "978-0-12-564134-0",
volume = "34",
series = "Methods in cell biology",
pages = "147--165",
editor = "Tartakoff, {Alan M.}",
booktitle = "Vectorial Pansport of Proteins into and across Membranes",

}

RIS

TY - CHAP

T1 - Preprotein Translocase of Escherichia coli: Solubilization, Purification, and Reconstitution of the Integral Membrane Subunits SecY/E

AU - Driessen, A. J. M.

AU - Brundage, L.

AU - Hendrick, Joseph P.

AU - Schiebel, E.

AU - Wickner, William

PY - 1991

Y1 - 1991

N2 - This chapter describes the procedures to solubilize and purify a functional secY/E protein that, upon reconstitution, supports an authentic translocation reaction of precursor proteins. The secY/E protein is purified from octylglucoside-extracted membranes by the ability of the reconstituted enzyme to stimulate the ATP-hydrolyzing activity of the purified secA protein in the presence of proOmpA. The chapter describes the reconstitution procedure and the various assays required to determine the activity of the reconstituted secY/E protein. Protocols used for the isolation and solubilization of E. coli inner membranes and the purification of the secY/E protein are described. The purified secY/E protein contains three major polypeptide species. These are identified by immunoblots with antisera to secY and secE and by N-terminal sequence analysis. The largest polypeptide of the purified protein reacts with antibodies to the secY N-terminus and migrates on SDS-PAGE with an apparent molecular mass of 29 kDa. The use of glycerol and phospholipids with octylglucoside has a dramatic effect on the solubility of the secY protein.

AB - This chapter describes the procedures to solubilize and purify a functional secY/E protein that, upon reconstitution, supports an authentic translocation reaction of precursor proteins. The secY/E protein is purified from octylglucoside-extracted membranes by the ability of the reconstituted enzyme to stimulate the ATP-hydrolyzing activity of the purified secA protein in the presence of proOmpA. The chapter describes the reconstitution procedure and the various assays required to determine the activity of the reconstituted secY/E protein. Protocols used for the isolation and solubilization of E. coli inner membranes and the purification of the secY/E protein are described. The purified secY/E protein contains three major polypeptide species. These are identified by immunoblots with antisera to secY and secE and by N-terminal sequence analysis. The largest polypeptide of the purified protein reacts with antibodies to the secY N-terminus and migrates on SDS-PAGE with an apparent molecular mass of 29 kDa. The use of glycerol and phospholipids with octylglucoside has a dramatic effect on the solubility of the secY protein.

KW - MALTOSE-BINDING PROTEIN

KW - PRECURSOR PROTEINS

KW - PLASMA-MEMBRANE

KW - TRIGGER FACTOR

KW - SECA PROTEIN

KW - PRO-OMPA

KW - FUNCTIONAL RECONSTITUTION

KW - CYTOPLASMIC MEMBRANE

KW - TRANSPORT-SYSTEM

KW - GENE-PRODUCT

U2 - 10.1016/S0091-679X(08)61679-9

DO - 10.1016/S0091-679X(08)61679-9

M3 - Chapter

SN - 978-0-12-564134-0

VL - 34

T3 - Methods in cell biology

SP - 147

EP - 165

BT - Vectorial Pansport of Proteins into and across Membranes

A2 - Tartakoff, Alan M.

ER -

ID: 62157032