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