Journal:

Article Title: Uptake of Pyocin S3 Occurs through the Outer Membrane Ferripyoverdine Type II Receptor of Pseudomonas aeruginosa

doi:

Figure Lengend Snippet: Plate showing the effects of different pyoverdines on the killing of P. aeruginosa ATCC 27853 by pyocin S3. To a lawn of P. aeruginosa ATCC 27853 on Luria-Bertani medium drops of purified pyocin S3 (20 μl, corresponding to 25 U ml−1, defined as the reciprocal of the highest pyocin dilution inhibiting bacterial growth) were deposited at three places in the middle. Next, on each side of the pyocin drop, 20 μl of the following purified pyoverdines (2 μmol) was added: that from PAO1 (top), that from ATCC 27853 (middle), and that from clinical isolate PA6 (bottom). Only the cognate pyoverdine (ATCC 27853), recognized by the pyoverdine receptor, competes with pyocin S3, which uses the same receptor, inhibiting the killing.

Article Snippet: The fact that mutants that do not produce the ferripyoverdine receptor are also defective in pyoverdine production has already been demonstrated ( 22 , 28 ), indicating that the biosynthesis of pyoverdine and the production of the receptor are coregulated. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Strain Pyocin S3 susceptibility Pyoverdine production a Growth on EDDHA with type II pyoverdine 59 Fe-pyoverdine incorporated (pmol) b Pb60 (WT) c + 100 + 264 Pb60R − 10 − 9 Pb60S + 0 + 236 ATCC 27853 (WT) + 100 + 257 ATCC 27853-5B − 0 − 7 ATCC 27853-6A − 26 − 6 ATCC 27853-6B − 0 − 7 ATCC 27853-9A1 − 0 − 13 ATCC 27853-14.2 − 7 − 0 ATCC 27853-16B1 − 9 − 0 ATCC 27853-16B2 − 9 − 0 Open in a separate window a Expressed as a percentage of the level produced by the wild type after 48 h of growth in CAA medium and after the absorbency at 400 nm was measured and normalized to growth at an absorbency of 600 nm ( 15 ). b Expressed in picomoles of 59 Fe incorporated per milligram (dry weight) of cells during a 20-min incubation.

Techniques: Purification

Journal:

Article Title: Uptake of Pyocin S3 Occurs through the Outer Membrane Ferripyoverdine Type II Receptor of Pseudomonas aeruginosa

doi:

Figure Lengend Snippet: Pyoverdine production, growth stimulation by pyoverdine, and uptake of ferripyoverdine of wild-type and pyocin S3-susceptible and -resistant mutants of strains Pb60 and ATCC 27853

Article Snippet: The fact that mutants that do not produce the ferripyoverdine receptor are also defective in pyoverdine production has already been demonstrated ( 22 , 28 ), indicating that the biosynthesis of pyoverdine and the production of the receptor are coregulated. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Strain Pyocin S3 susceptibility Pyoverdine production a Growth on EDDHA with type II pyoverdine 59 Fe-pyoverdine incorporated (pmol) b Pb60 (WT) c + 100 + 264 Pb60R − 10 − 9 Pb60S + 0 + 236 ATCC 27853 (WT) + 100 + 257 ATCC 27853-5B − 0 − 7 ATCC 27853-6A − 26 − 6 ATCC 27853-6B − 0 − 7 ATCC 27853-9A1 − 0 − 13 ATCC 27853-14.2 − 7 − 0 ATCC 27853-16B1 − 9 − 0 ATCC 27853-16B2 − 9 − 0 Open in a separate window a Expressed as a percentage of the level produced by the wild type after 48 h of growth in CAA medium and after the absorbency at 400 nm was measured and normalized to growth at an absorbency of 600 nm ( 15 ). b Expressed in picomoles of 59 Fe incorporated per milligram (dry weight) of cells during a 20-min incubation.

Techniques:

Journal:

Article Title: Uptake of Pyocin S3 Occurs through the Outer Membrane Ferripyoverdine Type II Receptor of Pseudomonas aeruginosa

doi:

Figure Lengend Snippet: Sodium dodecyl sulfate-polyacrylamide gel (7%) electrophoresis of 10-μg quantities of outer membrane proteins from P. aeruginosa ATCC 27853 grown in CAA medium (lane 1) and in CAA medium plus FeCl3 (50 μM) (lane 2), mutant ATCC 27853-5B (lane 3), mutant ATCC 27853-6A (lane 4), mutant ATCC 27853-9A1 (lane 5), and mutant ATCC 27853-16B1 (lane 6). Lane 7 contains the molecular mass markers (High molecular mass markers; Pharmacia-LKB, Uppsala, Sweden).

Article Snippet: The fact that mutants that do not produce the ferripyoverdine receptor are also defective in pyoverdine production has already been demonstrated ( 22 , 28 ), indicating that the biosynthesis of pyoverdine and the production of the receptor are coregulated. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Strain Pyocin S3 susceptibility Pyoverdine production a Growth on EDDHA with type II pyoverdine 59 Fe-pyoverdine incorporated (pmol) b Pb60 (WT) c + 100 + 264 Pb60R − 10 − 9 Pb60S + 0 + 236 ATCC 27853 (WT) + 100 + 257 ATCC 27853-5B − 0 − 7 ATCC 27853-6A − 26 − 6 ATCC 27853-6B − 0 − 7 ATCC 27853-9A1 − 0 − 13 ATCC 27853-14.2 − 7 − 0 ATCC 27853-16B1 − 9 − 0 ATCC 27853-16B2 − 9 − 0 Open in a separate window a Expressed as a percentage of the level produced by the wild type after 48 h of growth in CAA medium and after the absorbency at 400 nm was measured and normalized to growth at an absorbency of 600 nm ( 15 ). b Expressed in picomoles of 59 Fe incorporated per milligram (dry weight) of cells during a 20-min incubation.

Techniques: Electrophoresis, Mutagenesis