Solid-state NMR Study of the YadA Membrane-Anchor Domain in the Bacterial Outer Membrane.
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 E. coli in which it was expressed. YadA is an adhesin from the pathogen Yersinia enterocolitica 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 E. coli lipid environment by using (13) C-(13) C 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.
Max-Planck-Institute for Developmental Biology, Department 1, Tübingen (Germany).|Leibniz-Institut für Molekulare Pharmakologie FMP, Robert-Rössle-Str. 10, 13125 Berlin (Germany).
Shakeel A
SA
Shahid
Leibniz-Institut für Molekulare Pharmakologie FMP, Robert-Rössle-Str. 10, 13125 Berlin (Germany).
Madhu
M
Nagaraj
University of Oslo, Department of Biosciences, POBox 1066 Blindern, 0316 Oslo (Norway).
Nandini
N
Chauhan
Leibniz-Institut für Molekulare Pharmakologie FMP, Robert-Rössle-Str. 10, 13125 Berlin (Germany).
Trent W
TW
Franks
Unité de Bioinformatique Structurale, CNRS UMR3528, Institut Pasteur, Paris (France).
Benjamin
B
Bardiaux
Felix-Bernstein Institute for Mathematical Statistics, Georg-August-Universität Göttingen (Germany).|Max Planck Institute for Biophysical Chemistry, Göttingen (Germany).
Michael
M
Habeck
University of Oslo, Department of Biosciences, POBox 1066 Blindern, 0316 Oslo (Norway).
Marcella
M
Orwick-Rydmark
Max-Planck-Institute for Developmental Biology, Department 1, Tübingen (Germany). dirk.linke@ibv.uio.no.|University of Oslo, Department of Biosciences, POBox 1066 Blindern, 0316 Oslo (Norway). dirk.linke@ibv.uio.no.
Dirk
D
Linke
Leibniz-Institut für Molekulare Pharmakologie FMP, Robert-Rössle-Str. 10, 13125 Berlin (Germany). brossum@fmp-berlin.de.
Barth-J
BJ
van Rossum
D018829:Adhesins, Bacterial / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary; D015009:Yersinia Infections / Q000382:microbiology; D015008:Yersinia enterocolitica / Q000737:chemistry* / Q000235:genetics
D018829:Adhesins, Bacterial; D008055:Lipids; C064832:YadA protein, Yersinia
Yersinia adhesin A; autotransport; membrane proteins; solid-state NMR; structural biology