176.5
8.4
113.7
80.9
63.8
49.5
37.4
26.0
19.6
14.9
Bd1337
Bd2969 (L16P)
Bd3348
Bd0427Bd0286
Bd0633
Bd3340
Bd2994
Bd0427
Bd0459
Bd3037
Bd3340
Bd0010
Bd0160
Bd1015
Bd1475
Bd1290
Host-dependent membrane
proteins
Host-independent membrane
proteins
Fig 1
Fig 2
Av
era
ge
Ra
tio
(P
rote
in i
TR
AQ
va
lue
in
HD
10
0 s
tra
in/
HI1
00
str
ain
)
Fig 3
Av
era
ge
Ra
tio
(P
rote
in i
TR
AQ
va
lue
in
HD
10
0 s
tra
in/
HI1
00
str
ain
)
Fig 4
Fig 5
Av
era
ge
Ra
tio
(P
rote
in i
TR
AQ
va
lue
in
H
D1
00
str
ain
/ H
I10
0 s
tra
in)
Fig 6
Ave
rag
e R
atio
(P
rote
in iT
RA
Q v
alu
e in
HD
100
stra
in /
HI1
00 s
trai
n)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
Ave
rag
e R
atio
(P
rote
in iT
RA
Q v
alu
e in
HD
100
stra
in /
HI1
00 s
trai
n)
Fig 6
Membrane biogenesis Intracellular transportand secretion
Signal transduction Energy production Post-
translational modification
RT-PCR analysis of candidate membrane proteins( (Sockett Lab, UK)
Bd0112
Bd0427
Bd1337
Bd3180
Bd2611
M Ec HD 15’ 2h 4h3h HDDNA
Bd3000
1h HI
Intraperiplasmic stage
M: MarkerEc: E. coli S17-1 RNA only.HD : HD100 RNA onlyHI : HID2 RNA onlyHDDNA : HD100 genomic DNA
Fig 7
BBdellovibrio Grown on
Pseudomonas putida (HD100-P)
DHost Independent mutant
Bdellovibrio grown on host free medium.(HID2)
AHost Dependent
Bdellovibrio grown onEscherichia coli (HD100-E)
CBdellovibrio Grown on
Salmonella typhimurium(HD100-S)
HD100-E
HD100-P
HD100-S
HID2
Expt2
Experiment
1Experiment
2 Both datasets
Bdellovibrio peptides identified 1083 801
Number of Bdellovibrio membrane proteins identified 110 86 52
Proteins expressed over 1.5 fold in HD* 11 10 8
Proteins expressed over 1.5 fold in HI 16 18 9
7 923
Membrane proteins upregulated in HD strain
4
Expt1
Expt2
852 3458
Total proteins identified
Expt1
Expt2
Membrane proteins upregulated in HI strain
Expt1
9
* These proteins were upregulated in HD cells grown in atleast one host.
Summary of iTRAQ analysis of Bdellovibrio membrane proteins
A : Identified in one dataset ; B: Shows similar trend in 2DGE analysis ; C : Shows similar trend in SDS-PAGE analysis ; D : Shows similar trend in total protein iTRAQ analysis E: Not detected in iTRAQ analysis
Summary of candidate membrane proteinsORF Protein Name Predicted localisation
Membrane proteins expressed (1.5 fold or more) in Host-dependent strain
Bd0427 Major OMP Outer Membrane
Bd0459 A, C Hypothetical protein Outer Membrane
Bd1337C,D SWIB-domain containing proteins implicated in chromatin remodeling
Unknown
Bd3129 D Hypothetical protein Unknown
Bd2070 D Acetyl-CoA transferase Cytosolic
Bd3000 D, Bd3180 Hypothetical protein in Bdellovibrio only Outer Membrane
Bd3054 D Conserved hypothetical protein Unknown
Membrane proteins expressed (0.66 fold or less) in Host-dependent strain
Bd0063 A , Bd0260 B,E , Bd0293 A, D Hypothetical protein Outer Membrane
Bd0674 A, Bd1475 B,C , Bd1015 B,C, E Hypothetical protein in Bdellovibrio only Non-Cytosolic
Bd0999 D TolC protein Outer Membrane
Bd2611 Cytochrome c oxidase, cbb3-type, subunit I Inner Membrane
Bd2769 OmpA Outer Membrane
Bd1290 A,B,D PilA, Tfp pilus assembly protein Inner Membrane
Bd0220 D , Bd1703 Conserved hypothetical protein Unknown