Projection structure and oligomeric properties of a bacterial core protein translocase.
The major route for protein export or membrane integration in bacteria occurs via the Sec-dependent transport apparatus. The core complex in the inner membrane, consisting of SecYEG, forms a protein-conducting channel, while the ATPase SecA drives translocation of substrate across the membrane. The SecYEG complex from Escherichia coli was overexpressed, purified and crystallized in two dimensions. A 9 A projection structure was calculated using electron cryo-microscopy. The structure exhibits P12(1) symmetry, having two asymmetric units inverted with respect to one another in the unit cell. The map shows elements of secondary structure that appear to be transmembrane helices. The crystallized form of SecYEG is too small to comprise the translocation channel and does not contain a large pore seen in other studies. In detergent solution, the SecYEG complex displays an equilibrium between monomeric and tetrameric forms. Our results therefore indicate that, unlike other known channels, the SecYEG complex can exist as both an assembled channel and an unassembled smaller unit, suggesting that transitions between the two states occur during a functional cycle.
Collinson
I
I
Department of Cell Biology, Harvard Medical School/HHMI, Boston MA 02115, USA. ian_collinson@hms.harvard.edu
Breyton
C
C
Duong
F
F
Tziatzios
C
C
Schubert
D
D
Or
E
E
Rapoport
T
T
Kühlbrandt
W
W
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010458:Peptidyl Transferases / Q000737:chemistry*; D011233:Precipitin Tests; D000069816:SEC Translocation Channels; D012996:Solutions
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D009842:Oligopeptides; D000069816:SEC Translocation Channels; C065493:SecE protein, E coli; C057841:SecY protein, E coli; D012996:Solutions; D010458:Peptidyl Transferases