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A Specific Targeting Domain in Mature Exotoxin A Is Required for Its Extracellular Secretion From Pseudomonas Aeruginosa

A Specific Targeting Domain in Mature Exotoxin A Is Required for Its Extracellular Secretion From Pseudomonas Aeruginosa

H M Lu, S Lory

EMBO J. 1996 Jan 15;15(2):429-36.

PMID: 8617218

Abstract:

A number of Gram-negative bacteria, including Pseudomonas aeruginosa, actively secrete a subset of periplasmic proteins into their surrounding medium. The presence of a putative extracellular targeting signal within one such protein, exotoxin A, was investigated. A series of exotoxin A truncates, fused to beta-lactamase, was constructed. Hybrid proteins, which carry at their N- termini 120, 255, 355 or the entire 613 residues of the mature exotoxin A, were stable and were secreted into the extracellular medium. Hybrid proteins which carry residues 1-30 and 1-60 of the mature exotoxin A were unstable; however, they could be detected entirely within the cells after a short labeling period. A hybrid with beta-lactamase was constructed which carried only the N-terminal residues 1-3 and region 60-120 of exotoxin A. It was also secreted into the culture medium, suggesting that a specific 60 amino acid domain contains the necessary targeting information for translocation of exotoxin A across the outer membrane. The secretion of the hybrid proteins is independent of the passenger protein, since a similar exotoxin A-murine interleukin 4 hybrid protein was also secreted. The extracellular targeting signal between amino acids 60 and 120 is rich in anti-parallel beta-sheets. It has been shown previously to be involved in the interaction of the exotoxin A with the receptors of the eukaryotic cells. In the three- dimensional view, the targeting region is on the toxin surface where it is easily accessible to the components of the extracellular secretion machinery.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP91262952	Pseudomonas exotoxin A from Pseudomonas aeruginosa		91262-95-2	Price

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