Progesterone binding in a clinical isolate of Pseudomonas aeruginosa.
We have undertaken the characterization of progestin binding component(s) in the cytosol prepared from Pseudomonas aeruginosa isolated from an immunocompromised patient. Incubation of P. aeruginosa cytosol aliquots at 0 degrees C with 20 nM [3H]R5020 (a synthetic progestin) revealed the presence of saturable binding. The [3H]R5020 binding reached an equilibrium after 1 h at 0 degrees C and showed saturation at 30-50 nM with a Kd value of 7.7 nM. At 0 degrees C, beta-mercaptoethanol increased the [3H]R5020 binding by 20% but sodium molybdate had no effect. The [3H]R5020-macromolecular complex was stable for up to 4 h at 37 degrees C. Steroid binding specificity analysis revealed that [3H]R5020 binding could be eliminated in the presence of 2 microM progesterone, estradiol, or dihydrotestosterone but that the synthetic glucocorticoid, triamcinolone acetonide, did not compete. Postlabeling of the cytosol fractions obtained after 10-30% glycerol gradient analysis demonstrated association of the radioactivity with a molecule that sedimented as a 6-8 S protease-sensitive moiety which was unaltered in the presence of RNase or DNase. When cells were grown in the presence of 100 nM progesterone, a 50% inhibition in the number of resulting colonies was observed. In addition to its evolutionary significance, the presence of this steroid binding molecule suggests a potential in the endocrine manipulation in the treatment of infections caused by P. aeruginosa.
Mosier
S
S
Department of Biological Sciences, Oakland University, Rochester, Michigan 48309-4401.
Nakao
M
M
Herman
M
M
Walia
S K
SK
Rosenthal
N
N
Hurd
C
C
Moudgil
V K
VK
D001665:Binding Sites; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D011374:Progesterone / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000378:metabolism*; D014552:Urinary Tract Infections / Q000382:microbiology
D011374:Progesterone