Assay for Bacterial Membrane Protein Integration into Proteoliposomes

Overview

Journal Bio Protoc

Specialty Biology

Date 2021 Mar 4

PMID 33659299

Citations 4

Authors

Hanako Nishikawa

Masaru Sasaki

Ken-Ichi Nishiyama

Affiliations

Soon will be listed here.

Abstract

It is important to experimentally determine how membrane proteins are integrated into biomembranes to unveil the roles of the integration factors, and to understand the functions and structures of membrane proteins. We have developed a reconstitution system for membrane protein integration in using purified factors, in which the integration reaction is highly reproducible. This system enabled not only analysis of membrane-embedded factors including glycolipid MPIase, but also elucidation of the detailed mechanisms underlying membrane protein integration. Using the system, the integration of membrane proteins can be evaluated through a protease-protection assay. We report here how to prepare (proteo)liposomes and to determine the activities of membrane protein integration.

Citing Articles

Key contributions of a glycolipid to membrane protein integration.

Shimamoto K, Fujikawa K, Osawa T, Mori S, Nomura K, Nishiyama K Proc Jpn Acad Ser B Phys Biol Sci. 2024; 100(7):387-413.

PMID: 39085064 PMC: 11413397. DOI: 10.2183/pjab.100.026.

Roles of a Glycolipid MPIase in Sec-Independent Membrane Protein Insertion.

Nomura K, Mori S, Shimamoto K Membranes (Basel). 2024; 14(2).

PMID: 38392675 PMC: 10890265. DOI: 10.3390/membranes14020048.

Novel Proteoliposome-Based Vaccine against : A Potential New Tool for the Control of Bovine Mastitis.

Quiroga J, Vidal S, Siel D, Caruffo M, Valdes A, Cabrera G Animals (Basel). 2022; 12(19).

PMID: 36230275 PMC: 9558995. DOI: 10.3390/ani12192533.

Interplay between MPIase, YidC, and PMF during Sec-independent insertion of membrane proteins.

Endo Y, Shimizu Y, Nishikawa H, Sawasato K, Nishiyama K Life Sci Alliance. 2021; 5(1).

PMID: 34642230 PMC: 8548208. DOI: 10.26508/lsa.202101162.

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