Secretion of the STA3 heat-stable enterotoxin of Escherichia coli: extracellular delivery of Pro-STA is accomplished by either Pro or STA.
The methanol-soluble, heat-stable enterotoxin of Escherichia coli is a protease-resistant extracellular peptide which is synthesized as a 72-amino-acid precursor Pre-Pro-STA3. The specific roles of Pre (19 amino acids), Pro (34 amino acids) and STA3 (19 amino acids) in the secretion process were studied by functionally deleting each of the three domains. Deletion of the Pre signal sequence resulted in a short-lived cell-associated molecule with an M(r) equivalent to that of Pro-STA3. Deletion of Pro (i.e., Pre-STA3) resulted in the rapid extracellular accumulation of STA3; the periplasmic intermediate found in the secretion of the wild-type toxin was undetected. Deletion of the STA3 domain resulted in a cell-associated Pre-Pro peptide; with time this form converted to periplasmic Pro which later became extracellular. When DNA encoding either STA3, by itself, or Pro-STA3 (lacking the signal peptide) was expressed, these peptides were degraded intracellularly, with no periplasmic or extracellular forms detected. The results presented demonstrate that the signal peptide (Pre) is essential even for the export of small peptides to the periplasm, and that its absence causes the STA3 domain to become susceptible to intracellular proteases. The rapid degradation of intracellular STA3 indicates that its proteolytic resistance is acquired in a compartment other than the cytoplasm. The results also show that after the Pre domain is proteolytically cleaved from Pre-STA3 and Pre-Pro, the STA3 and Pro peptides can exit to the culture supernatant.(ABSTRACT TRUNCATED AT 250 WORDS)
Yang
Y
Y
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235-9048.
Gao
Z
Z
Guzmán-Verduzco
L M
LM
Tachias
K
K
Kupersztoch
Y M
YM
D000595:Amino Acid Sequence; D001427:Bacterial Toxins / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D004768:Enterotoxins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000378:metabolism; D021382:Protein Sorting Signals / Q000378:metabolism; D017384:Sequence Deletion; D013329:Structure-Activity Relationship
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; C034151:heat stable toxin (E coli)