Nature - USA (2020-08-20)

a

M212AM212EM212KT213VT213DR215AR216AR216ER216DM212A/

T213V/

R216A

vector WT WT vector D268AT302VR451A

b

+WT PbgA

+D268A

+T302A

+R451A

+WT PbgA

+T213V

+T213D

+M212E

+M212K

+M212A

+WT PbgA

+MTR-AVA +R216D

+R216A

+R216E

+WT PbgA

+R215A

0 10 -3 10 -2 10 -1 100 101 102

0.03

0.1

0.5

OD

600

0 10 -3 10 -2 10 -1 100 101 102

0.03

0.1

0.5

OD

600

0 10 -3 10 -2 10 -1 100 101 102

0.03

0.1

0.5

OD

600

0 10 -3 10 -2 10 -1 100 101 102

0.03

0.1

0.5

OD

600

Extended Data Fig. 6 | PbgA mutants and outer membrane permeability.
a, All UPEC-ΔpbgA bacteria tested in the rifampicin sensitivity assay were
probed by western blot analysis to confirm PbgA–Flag expression. GroEL was
assessed as a loading control. Representative blots for n = 3 or more
experiments are shown. b, Outer membrane permeability of UPEC ΔpbgA
strains with pBADpbgA plasmids expressing wild-type or mutant pbgA

assessed by rifampicin sensitivity, where MTR-AVA is the M212A/T213V/R216A

PbgA triple mutant. Data are representative and presented as mean ± s.d. for

n = 3 or more independent cultures. Note, see Extended Data Fig. 2f for a view of

the salt-bridge interaction between R215 (IFD) and a conserved TMD acidic

residue, D192.