Structure analysis of the membrane protein TatC(d) from the Tat system of B. subtilis by circular dichroism.
The twin arginine translocation (Tat) system can transport fully folded proteins, including their cofactors, across bacterial and thylakoid membranes. The Tat system of Bacillus subtilis that serves to export the phosphodiesterase (PhoD) consists of only two membrane proteins, TatA(d) and TatC(d). The larger component TatC(d) has a molecular weight of 28 kDa and several membrane-spanning segments. This protein has been expressed in Escherichia coli and purified in sufficient amounts for structure analysis by circular dichroism (CD) and NMR spectroscopy. TatC(d) was reconstituted in detergent micelles and in lipid bilayers for CD analysis in solution and in macroscopically oriented samples, to examine the stability of the protein. Suitable protocols and model membrane systems have been established, by which TatC(d) maintains the level of helicity close to theoretically predicted, and its transmembrane alignment could been verified.
Nolandt
Olga V
OV
Karlsruhe Institute of Technology, Institut für Biologische Grenzflächen , Forschungszentrum Karlsruhe, D-76021 Karlsruhe, Germany.
Walther
Torsten H
TH
Roth
Siegmar
S
Bürck
Jochen
J
Ulrich
Anne S
AS
D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology*; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D008051:Lipid Bilayers / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D008051:Lipid Bilayers; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; D001120:Arginine