Topology and subcellular localization of FtsH protein in Escherichia coli.
FtsH protein in Escherichia coli is an essential protein of 70.7 kDa (644 amino acid residues) with a putative ATP-binding sequence. Western blots (immunoblots) of proteins from fractionated cell extracts and immunoelectron microscopy of the FtsH-overproducing strain showed exclusive localization of the FtsH protein in the cytoplasmic membrane. Most of the FtsH-specific labeling with gold particles was observed in the cytoplasmic membrane and the adjacent cytoplasm; much less was observed in the outer membrane and in the bulk cytoplasm. Genetic analysis by TnphoA insertions into ftsH revealed that the 25- to 95-amino-acid region, which is flanked by two hydrophobic stretchs, protrudes into the periplasmic space. From these results, we concluded that FtsH protein is an integral cytoplasmic membrane protein spanning the membrane twice and that it has a large cytoplasmic carboxy-terminal part with a putative ATP-binding domain. The average number of FtsH molecules per cell was estimated to be approximately 400.
Tomoyasu
T
T
Department of Molecular Cell Biology, Kumamoto University School of Medicine, Japan.
Yamanaka
K
K
Murata
K
K
Suzaki
T
T
Bouloc
P
P
Kato
A
A
Niki
H
H
Hiraga
S
S
Ogura
T
T
D049069:ATP-Dependent Proteases; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000737:chemistry* / Q000648:ultrastructure; D029968:Escherichia coli Proteins; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D015183:Restriction Mapping
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000255:Adenosine Triphosphate; D049069:ATP-Dependent Proteases; C513383:FtsH protein, E coli