2. P4
Nascent Mature
OMP
Assembled
Porin
Sec
Cyt
BamB
BamC
BamD
BamA
BamE
Precursor
OMP
P5
P3
P2
P1
Chaperones
(SurA, Skp)
Per
Gram-negative Bacteria
OM
IM

3. What is the minimal amount of BAM A Complex you can
create in E coli?
How can E coli compensate for loss of the other BAM
proteins?

4. BamABCDE BamACD (Conditional Lethal) BamA*CD
ΔBamBE
BamA*CD
ΔBamC
BamA*D (Conditonal Lethal) BamA*D
X*
VRGY(F)
R541 (E780)
Q440 (N631)
T285 (F474) G444 (G635)
Q458 (G649)
(A652)
L6 loop

5. Retro-evolution Re-evolution
OmpACF 1 .83 .81 .60 .47
OmpA 1 .78 .83 .51 .60
OmpC 1 1 .97 .89 .79
OmpF 1 .82 .84 .34 .17
OmpACF 1 .92 1.28 .95 .1.47
OmpA 1 .77 1 .83 1.37
OmpC 1 1 1.39 1.1 2
OmpF 1 .50 1.14 .87 .11
OMPs OMPs

6. My story begins with 6 Mutants…..
1D
2C
3B
4A
4C
4D
λ
λ

10. 0
0.5
1
1.5
2
2.5
3
3.5
4
1827 126 1D 2C 3B 4A 4C 4D
Omp C
Omp
C
-0.4
-0.2
0
0.2
0.4
0.6
0.8
1
1.2
1827 126 1D 2C 3B 4A 4C 4D
Omp F
Omp F
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1827 126 1D 2C 3B 4A 4C 4D
Omp A
Omp A

11. 0
5
10
15
20
25
30
1827 126 1D 2C 3B 4A 4C 4D
NOVOBIOCIN

12. 0
20
40
60
80
100
120
140
160
0 50 100 150 200 250 300 350
1827 Av
126 Av
1D Av
4A Av
Slopes:
1827: -3.0063(a.u.)/(s) ± 0.393252
126: -2.07445 (a.u.)/(s) ± 0.386454
1D: -4.69265 (a.u.)/(s) ± 0.795373
4A: -3.77925 (a.u.)/(s) ± 0.451426

14. 0
2
4
6
8
10
12
1827 126 1D 2C 3B 4A 4C 4D
IntensityINT*mm2
Strains
BAM A Intensity Analysis
STRAINS BAM A Control
1827 8.663435 5.967747
126 9.712198 5.6763
1D 8.338653 5.314919
2C 7.181835 5.497507
3B 6.618693 5.006862
4A 7.571512 4.823855
4C 7.863744 5.502401
4D 8.292057 4.677809