176.5 8.4 113.7 80.9 63.8 49.5 37.4 26.0 19.6 14.9 Bd1337 Bd2969 (L16P) Bd3348 Bd0427 Bd0286 Bd0633...

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