The transition state for folding of an outer membrane protein.


Abstract

Inspired by the seminal work of Anfinsen, investigations of the folding of small water-soluble proteins have culminated in detailed insights into how these molecules attain and stabilize their native folds. In contrast, despite their overwhelming importance in biology, progress in understanding the folding and stability of membrane proteins remains relatively limited. Here we use mutational analysis to describe the transition state involved in the reversible folding of the beta-barrel membrane protein PhoPQ-activated gene P (PagP) from a highly disordered state in 10 M urea to a native protein embedded in a lipid bilayer. Analysis of the equilibrium stability and unfolding kinetics of 19 variants that span all eight beta-strands of this 163-residue protein revealed that the transition-state structure is a highly polarized, partly formed beta-barrel. The results provide unique and detailed insights into the transition-state structure for beta-barrel membrane protein folding into a lipid bilayer and are consistent with a model for outer membrane protein folding via a tilted insertion mechanism. Study holds ProTherm entries: 25469, 25470, 25471, 25472, 25473, 25474, 25475, 25476, 25477, 25478, 25479, 25480, 25481, 25482, 25483, 25484, 25485, 25486, 25487, 25488 Extra Details: beta barrel; membrane protein; PagP; phi-value analysis; protein folding

Submission Details

ID: yyMkht8D3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:55 p.m.

Version: 1

Publication Details
Huysmans GH;Baldwin SA;Brockwell DJ;Radford SE,Proc. Natl. Acad. Sci. U.S.A. (2010) The transition state for folding of an outer membrane protein. PMID:20133664
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1MM5 2002-09-13 Solution NMR structure of the outer membrane enzyme PagP in OG micelles
1MM4 2002-09-13 Solution NMR structure of the outer membrane enzyme PagP in DPC micelles
3GP6 2010-06-23 1.4 Crystal structure of PagP in SDS/MPD
1THQ 2004-08-10 1.9 Crystal Structure of Outer Membrane Enzyme PagP

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.1 Lipid A palmitoyltransferase PagP Q1RET8 PAGP_ECOUT
98.9 Lipid A palmitoyltransferase PagP Q324S0 PAGP_SHIBS
99.5 Lipid A palmitoyltransferase PagP Q83LW9 PAGP_SHIFL
99.5 Lipid A palmitoyltransferase PagP Q0T6N3 PAGP_SHIF8
99.5 Lipid A palmitoyltransferase PagP D2AA19 PAGP_SHIF2
99.5 Lipid A palmitoyltransferase PagP E0J1Q4 PAGP_ECOLW
99.5 Lipid A palmitoyltransferase PagP E8Y6Y6 PAGP_ECOKO
98.8 Lipid A palmitoyltransferase PagP Q8FJZ7 PAGP_ECOL6
98.8 Lipid A palmitoyltransferase PagP Q0TK50 PAGP_ECOL5
99.5 Lipid A palmitoyltransferase PagP Q8XBR9 PAGP_ECO57
100.0 Lipid A palmitoyltransferase PagP P37001 PAGP_ECOLI