Glutathione transferase in bacteria: subunit composition and antigenic characterization.
The presence of glutathione transferase (GST; EC 2.5.1.18) in Escherichia coli ATCC 25922, E. coli ATCC 25422, Proteus vulgaris ATCC 8427, Pseudomonas aeruginosa ATCC 27853, Klebsiella oxytoca CIP 666, K. oxytoca AF 101, Enterobacter cloacae CIP 6085, Serratia marcescens CIP 6755, and Proteus mirabilis AF 2924 was investigated. Using 1-chloro-2,4-dinitrobenzene as substrate, GST activity was found in the glutathione-(GSH-)affinity-purified fraction of all strains tested. SDS-PAGE analysis of GSH-affinity-purified enzyme indicated that the GSTs of all these bacteria are dimers of two identical subunits of Mr about 22,500. Rabbit antiserum directed against the major isoenzyme present in Proteus mirabilis AF 2924, Pm-GST-6.0, was used to investigate the antigenic properties of bacterial GSTs. Western blot analysis indicated that a GST antigenically identical to Pm-GST-6.0 is present in Enterobacter cloacae CIP 6085, Escherichia coli ATCC 25422 and Proteus vulgaris ATCC 8427, but absent in Escherichia coli ATCC 25922, Klebsiella oxytoca CIP 666, K. oxytoca AF 101 and Serratia marcescens CIP 6755. The presence of Pm-GST-6.0, but not mammalian GST, increased the MIC values of amikacin, ampicillin, cefotaxime, cephalothin and nalidixic acid for E. coli ATCC 25922. It is suggested that bacterial GST may represent a defense against the effects of antibiotics.
Piccolomini
R
R
Istituto di Medicina Sperimentale, Cattedra di Microbiologia, Chieti, Italy.
Di Ilio
C
C
Aceto
A
A
Allocati
N
N
Faraone
A
A
Cellini
L
L
Ravagnan
G
G
Federici
G
G
D000900:Anti-Bacterial Agents / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000942:Antigens, Bacterial / Q000302:isolation & purification; D001419:Bacteria / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D003429:Cross Reactions; D005982:Glutathione Transferase / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D008970:Molecular Weight; D011487:Protein Conformation
D000900:Anti-Bacterial Agents; D000942:Antigens, Bacterial; D007527:Isoenzymes; D005982:Glutathione Transferase