title: Heat Modifiability of Outer Membrane Proteins from Gram-Negative Bacteria.
abstract: β-barrel membrane proteins are somewhat unique in that their folding states can be monitored using semi-native SDS-PAGE methods to determine if they are folded properly or not. This property, which is commonly referred to as heat modifiability, has been used for many years on both purified protein and on whole cells to monitor folded states of proteins of interest. Additionally, heat modifiability assays have proven indispensable in studying the BAM complex and its role in folding and inserting β-barrel membrane proteins into the outer membrane. Here, we describe the protocol our lab uses for performing the heat modifiability assay in our studies on outer membrane proteins.
authors.affiliation: Department of Biological Sciences, Markey Center for Structural Biology, Purdue University, West Lafayette, IN, 47907, USA. nnoinaj@purdue.edu.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Noinaj
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892, USA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Kuszak
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892, USA.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Buchanan
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D006090:Gram-Negative Bacteria; D006358:Hot Temperature; D017510:Protein Folding
chemical_list: D001425:Bacterial Outer Membrane Proteins
keywords: BAM complex; BamA; Heat modifiability; OMP; Outer membrane protein; Protein folding; β-barrel membrane protein