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PMID: 8418052 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Purification and characterization of exoenzyme S from Pseudomonas aeruginosa 388.

Infection and immunity ·Vol. 61 ·No. 1 ·1993-01-00 ·Pages 307-13

Kulich SM, Frank DW, Barbieri JT

Abstract

Exoenzyme S was purified > 1,500-fold from the culture supernatant fluid of Pseudomonas aeruginosa 388 at high yield without utilization of solvents or detergents. Two proteins, with apparent molecular sizes of 53 and 49 kDa, cofractionated with exoenzyme S activity. Rabbit anti-49-kDa-protein immunoglobulin G was prepared by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis-purified 49-kDa protein as immunogen. Anti-49-kDa-protein IgG inhibited the ADP-ribosyltransferase activity of purified exoenzyme S in a dose-dependent manner, which indicated a role for the 49-kDa protein in the ADP-ribosylation reaction. Analysis by ultrafiltration showed that exoenzyme S activity and the 53- and 49-kDa proteins cofractionated and that exoenzyme S was apparently > 300 kDa in size. Urea (8 M) and 1.0% Triton X-100 reversibly decreased the apparent molecular sizes of exoenzyme S activity and the 53- and 49-kDa proteins to between 30 and 100 kDa.

MeSH Terms
ADP Ribose Transferases Amino Acids/analysis Animals Antibody Specificity Bacterial Toxins Blotting, Western Chromatography, Ion Exchange Dose-Response Relationship, Immunologic Electrophoresis, Polyacrylamide Gel Immunoglobulin G/pharmacology Mice Octoxynol Poly(ADP-ribose) Polymerases/chemistry,isolation & purification,metabolism Polyethylene Glycols/pharmacology Pseudomonas aeruginosa/enzymology Ultrafiltration Urea/pharmacology
Chemicals
Amino Acids Bacterial Toxins Immunoglobulin G Polyethylene Glycols Urea Octoxynol ADP Ribose Transferases Poly(ADP-ribose) Polymerases exoenzyme S
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kulich S M
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
Frank D W
Barbieri J T
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.61.1.307-313.1993
Published
1993-01-00
Pages
307-13
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC302720
Subset
IM
Grants
NIAID NIH HHS · AI31665 · United States
NCRR NIH HHS · RR-03326 · United States
External Links
PubMed source DOI: 10.1128/iai.61.1.307-313.1993 PMC Full Text View journal details
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