Nature - USA (2020-10-15)

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Free

CDN

[α^32 P] CDN:

STING:

c-di-AMPc-di-GMP3',3'-cGAMP

CgSTING ΔTIR

2',3'-cGAMPc-di-AMPc-di-GMP3',3'-cGAMP2',3'-cGAMP

No protein

STING–CDN

Complex

Well

f

g

h

10 −4 10 −3 10 −2 10 −1 100 101 102 103

0.0

0.4

0.8

1.2

[ReSTING ΔTM] (μM)

Fraction boun

d 3',3'-cGAMP

c-di-GMP

10 −4 10 −3 10 −2 10 −1 100 101 102 103

0.0

0.4

0.8

1.2

[SfSTING] (μM)

Fraction boun

d

c-di-GMP

3',3 '-cGAMP

AaSTING (TIR)

c-di-AMPc-di-GMP3',3'-cGAMP2',3'-cGAMP

Free

CDN

[α^32 P] CDN:

STING:LbSTING ΔTIR

c-di-AMPc-di-GMP3',3'

-cGAMP

2',3'-cGAMP

Well

STING–CDN

Complex

Free

c-di-GMP

STING:— SfSTING (TIR

)

Aa

STING (TIR

)

Re

STING ΔT

M

Nd

STING ΔTIR

Cg

ST

ING ΔTIR

FsSTING ΔT LbSTING ΔTIR

M

Fd

STING ΔTIR

Well

STING–CDN

Complex

STING:—

CgSTING ΔTIR

Free

c-di-GMP

STING:—

CgSTING ΔTIR

Free

3',3'-cGAMP

10 −4 10 −3 10 −2 10 −1 100 101 102 103

0.0

0.4

0.8

1.2

[CgSTING ΔTIR] (μM)

Fraction bound

c-di-GMP

Well Well 3',3'-cGAMP

STING–CDN

Complex

STING–CDN

Complex

STING:—

SfSTING (TIR)

Free

c-di-GMP

Well

STING–CDN

Complex

STING:—

SfSTING (TIR)

Free

3',3'-cGAMP

Well

STING–CDN

Complex

STING:—

ReSTING ΔTM

Free

c-di-GMP

Well

STING–CDN

Complex

STING:—

ReSTING ΔTM

Free

3',3'-cGAMP

Well

STING–CDN

Complex

Free

3',3'-c-UMP–AMP

STING:— SfST

ING (TIR

)

Aa

STING (TIR

)

Re

STING ΔTM

Nd

STIN

G ΔTIR

Cg

STING ΔTIR

FsSTING ΔT LbSTING ΔTIR

M

Fd

STING ΔTIR

Well

STING–CDN

Complex

de

SfSTING (TIR)

ReSTING ΔTM

CgSTING ΔTIR

c-di-GMP

Kd (μM)

3',3'-cGAMP

Kd (μM)

0.29 0.69

0.75 36.31

0.83 11.74

STING construct

Extended Data Fig. 4 | See next page for caption.