Rapid Exploration of the Folding Topology of Helical Membrane Proteins Using Paramagnetic Perturbation

Overview

Journal Protein Sci

Specialty Biochemistry

Date 2010 Oct 15

PMID 20945360

Citations 2

Authors

Kwon Joo Yeo

Hye-Yeon Kim

Young Pil Kim

Eunha Hwang

Myung Hee Kim

Chaejoon Cheong

Senyon Choe

Young Ho Jeon

Affiliations

Soon will be listed here.

Abstract

An understanding of the folding states of α-helical membrane proteins in detergent systems is important for functional and structural studies of these proteins. Here, we present a rapid and simple method for identification of the folding topology and assembly of transmembrane helices using paramagnetic perturbation in nuclear magnetic resonance spectroscopy. By monitoring the perturbation of signals from glycine residues located at specific sites, the folding topology and the assembly of transmembrane helices of membrane proteins were easily identified without time-consuming backbone assignment. This method is validated with Mistic (membrane-integrating sequence for translation of integral membrane protein constructs) of known structure as a reference protein. The folding topologies of two bacterial histidine kinase membrane proteins (SCO3062 and YbdK) were investigated by this method in dodecyl phosphocholine (DPC) micelles. Combing with analytical ultracentrifugation, we identified that the transmembrane domain of YbdK is present as a parallel dimer in DPC micelle. In contrast, the interaction of transmembrane domain of SCO3062 is not maintained in DPC micelle due to disruption of native structure of the periplasmic domain by DPC micelle.

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