Tags

Type your tag names separated by a space and hit enter

Development of a functional in vitro integration system for an integral membrane protein, SecG.
Biochem Biophys Res Commun 2009; 390(3):920-4BB

Abstract

A functional in vitro integration system for an integral membrane protein, SecG, comprising an efficient translation system supplemented with inverted membrane vesicles (IMV) was developed. When SecG was synthesized in the presence of IMV prepared from a DeltasecG strain (DeltaSecG IMV), the synthesized SecG was recovered with the IMV. A population of SecG was resistant to urea extraction, indicating that the synthesized SecG was integrated into DeltaSecG IMV. Addition of signal recognition particle and its receptor (SRP) and SecA caused an increase in the amount of the urea-resistant form of SecG. When IMV into which SecG had been integrated were subjected to the translocation assay, the translocation activity was found to be significantly stimulated compared with for DeltaSecG IMV. Moreover, when SRP and SecA had been supplemented, the translocation activity nearly recovered to the level in IMV prepared from the wild type strain. These results indicate that the in vitro synthesized SecG could be functionally integrated into DeltaSecG IMV with the help of SRP and SecA. We also present evidence that the membrane targeting and integration of SecG is stimulated by externally added SecA and SecG itself.

Authors+Show Affiliations

Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo 113-0032, Japan. unishiy@mail.ecc.u-tokyo.ac.jpNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19853580

Citation

Nishiyama, Ken-ichi, and Hajime Tokuda. "Development of a Functional in Vitro Integration System for an Integral Membrane Protein, SecG." Biochemical and Biophysical Research Communications, vol. 390, no. 3, 2009, pp. 920-4.
Nishiyama K, Tokuda H. Development of a functional in vitro integration system for an integral membrane protein, SecG. Biochem Biophys Res Commun. 2009;390(3):920-4.
Nishiyama, K., & Tokuda, H. (2009). Development of a functional in vitro integration system for an integral membrane protein, SecG. Biochemical and Biophysical Research Communications, 390(3), pp. 920-4. doi:10.1016/j.bbrc.2009.10.078.
Nishiyama K, Tokuda H. Development of a Functional in Vitro Integration System for an Integral Membrane Protein, SecG. Biochem Biophys Res Commun. 2009 Dec 18;390(3):920-4. PubMed PMID: 19853580.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a functional in vitro integration system for an integral membrane protein, SecG. AU - Nishiyama,Ken-ichi, AU - Tokuda,Hajime, Y1 - 2009/10/22/ PY - 2009/10/09/received PY - 2009/10/15/accepted PY - 2009/10/27/entrez PY - 2009/10/27/pubmed PY - 2009/12/30/medline SP - 920 EP - 4 JF - Biochemical and biophysical research communications JO - Biochem. Biophys. Res. Commun. VL - 390 IS - 3 N2 - A functional in vitro integration system for an integral membrane protein, SecG, comprising an efficient translation system supplemented with inverted membrane vesicles (IMV) was developed. When SecG was synthesized in the presence of IMV prepared from a DeltasecG strain (DeltaSecG IMV), the synthesized SecG was recovered with the IMV. A population of SecG was resistant to urea extraction, indicating that the synthesized SecG was integrated into DeltaSecG IMV. Addition of signal recognition particle and its receptor (SRP) and SecA caused an increase in the amount of the urea-resistant form of SecG. When IMV into which SecG had been integrated were subjected to the translocation assay, the translocation activity was found to be significantly stimulated compared with for DeltaSecG IMV. Moreover, when SRP and SecA had been supplemented, the translocation activity nearly recovered to the level in IMV prepared from the wild type strain. These results indicate that the in vitro synthesized SecG could be functionally integrated into DeltaSecG IMV with the help of SRP and SecA. We also present evidence that the membrane targeting and integration of SecG is stimulated by externally added SecA and SecG itself. SN - 1090-2104 UR - https://www.unboundmedicine.com/medline/citation/19853580/Development_of_a_functional_in_vitro_integration_system_for_an_integral_membrane_protein_SecG_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(09)02069-5 DB - PRIME DP - Unbound Medicine ER -