A fast way to track functional OmpF reconstitution in liposomes: Escherichia coli total lipid extract

• Tipo: Artigo em Revista Científica Internacional
• Data: 2015

Autores

Lopes, SC (Autor) Outra; Ferreira, M (Autor) Outra; Sousa, CF (Autor) Outra; Gameiro, P (Autor) FCUP

Revista

Título: Analytical Biochemistry

Vol. 479

Páginas: 54-59

ISSN: 0003-2697

Editora: Elsevier

Indexação

ISI Web of Knowledge - 3 Citações

Scopus - 3 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Química

Outras Informações

ID Authenticus: P-00A-CW0

DOI: 10.1016/j.ab.2015.03.026

Abstract (EN): A major requirement to perform structural studies with membrane proteins is to define efficient reconstitution protocols that ensure a high incorporation degree and protein directionality and topology that mimics its in vivo conditions. For this kind of studies, protein reconstitution in membrane systems via a detergent-mediated pathway is usually successfully adopted because detergents are generally used in the initial isolation and purification of membrane proteins. This study reports OmpF reconstitution in preformed Escherichia coli liposomes followed by detection of its insertion by analyzing modifications on membrane structure by two different techniques: steady-state fluorescence anisotropy and dynamic light scattering. Another important issue is protein directionality. For OmpF, it is known that interaction with polyamines promotes channel blockage. In this work, the spermine-OmpF interaction was evaluated using surface plasmon resonance, and protein directionality was confirmed.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 6