Solid‐state NMR Study of the YadA Membrane‐Anchor Domain in the Bacterial Outer Membrane. Issue 43 (31st August 2015)

• Record Type: Journal Article
• Authors: Shahid, Shakeel A.; Nagaraj, Madhu; Chauhan, Nandini; Franks, Trent W.; Bardiaux, Benjamin; Habeck, Michael; Orwick‐Rydmark, Marcella; Linke, Dirk; van Rossum, Barth‐J.
• Is Part Of: Angewandte Chemie international edition. Volume 54:Issue 43(2015)
• Page Start: 12602
• Page End: 12606
• Publication Date: 2015-08-31
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201505506
• Languages: English
• ISSNs: 1433-7851

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>MAS‐NMR was used to study the structure and dynamics at ambient temperatures of the membrane‐anchor domain of YadA (YadA‐M) in a pellet of the outer membrane of <italic>E</italic>. <italic>coli</italic> in which it was expressed. YadA is an adhesin from the pathogen <italic>Yersinia enterocolitica</italic> that is involved in interactions with the host cell, and it is a model protein for studying the autotransport process. Existing assignments were sucessfully transferred to a large part of the YadA‐M protein in the <italic>E</italic>. <italic>coli</italic> lipid environment by using ¹³C‐¹³C DARR and PDSD spectra at different mixing times. The chemical shifts in most regions of YadA‐M are unchanged relative to those in microcrystalline YadA‐M preparations from which a structure has previously been solved, including the ASSA region that is proposed to be involved in transition‐state hairpin formation for transport of the soluble domain. Comparisons of the dynamics between the microcrystalline and membrane‐embedded samples indicate greater flexibility of the ASSA region in the outer‐membrane preparation at physiological temperatures. This study will pave the way towards MAS‐NMR structure determination of membrane proteins, and a better understanding of functionally important dynamic residues in native membrane environments.</p> </abstract>