The membrane binding C-terminus of protein A from Staphylococcus aureus affects its cellular localization and causes structural deformation when expressed in Escherichia coli.
Protein A from Staphylococcus aureus is a powerful diagnostic reagent and has several uses in human disease therapy. Expression in non-pathogenic Escherichia coli containing recombinant plasmids coding for this protein has increased its availability, but can reduce the stability of the plasmid-bearing host. By employing immune electron microscopy, we have determined that E. coli containing stable plasmids coding for a truncated version of protein A, without the membrane binding site, secrete this protein through the cytoplasmic membrane and into the periplasmic space, where it accumulates. E. coli containing unstable plasmids, however, which code for the complete protein including the membrane-binding site, target the protein into the cytoplasmic membrane. This accumulation of protein A in the E. coli cytoplasmic membrane inhibits the formation of septa between dividing cells and results in aberrant elongated, multi-chromosomal forms.
Warnes
A
A
PHLS Centre for Applied Microbiology and Research, Salisbury, UK.
Brown
M R
MR
Fooks
A R
AR
Shuttleworth
H
H
Dowsett
A B
AB
Melling
J
J
Stephenson
J R
JR
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000235:genetics*
D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A