Nature - USA (2020-08-20)

Article

a

c

LPS

CL

PG

PE

1 0 1 0 1 0 1 0

response (nm)

LABWT LABWT+ LABT213D

0

150

100

50

25

10

KD (LPS)

LABWT 73±13μM

LABWT+ 52±11μM

LABT213D n.d.

n.d.

LABWT SYPMTARRFLEKHGLLD

LABWT+ SYPMTARRFLEKWGLLR

LABT213D SYPMDARRFLEKWGLLR

SYPMTARRFLE

peptide sequence

no compound

LABv2.1 1x MIC

LABv2.1 0.5x MIC

LABv2.1 2x MIC

polymyxin B 1.25 μg/ml

LABWT

LABWT+

LABv2.0

LABv2.1_minus1_Dap212=Mat213

LABv2.1

LABv2.1_Dap213Thr

LABv2.1_Dap21Arg

b

d

102

103

104

105

106

107

108

CFU/mL

050100 150

time (min)

2.0

1.5

1.0

0.5

0

OD

405

PBS water

Triton X-100

% RBC lysis (vs. water)

0100 200300 400500

0

100

50

Extended Data Fig. 7 | Characterization of PbgA-derived, synthetic LAB
peptides. a, Biotinylated LAB peptides were captured and interferometry
measurements measured upon presenting peptides to different
concentrations of detergent solubilized lipids (LPS, phosphatidylethanolamine,
phosphatidylglycerol and cardiolipin). Three independent experiments were
performed and data shown are representative. b, CFUs of E. coli ATCC 25922
measured over time with LABv2.1 and polymyxin B. Data are mean ± s.d. for n = 3

independent cultures. c, A red blood cell (RBC) lysis assay evaluated after 18 h

in the presence of indicated compounds (Methods). Data are mean ± s.d. (n = 3)

for each compound tested. d, A RBC lysis assay comparing LABv2.1 precursors

(LABWT, LABW T+, LABv2.0) and LABv2.1 analogues designed, based on the

LPS–PbgA crystal structure, to disrupt specific interactions of lipid A

(LABv2.1_Dap213Thr, LABv2.1_Dap213Arg, LABv2.1_Dap212-Met213). Data are mean ± s.d. for n = 3

independent assay of each compound at each concentration.