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Table 2 Ability of Gα16/z chimeras to stimulate IP3 production in HEK 293 cells.

From: 16 interacts with tetratricopeptide repeat 1 (TPR1) through its β3 region to activate Ras independently of phospholipase Cβ signaling

Construct

Intact PLCβ Domain

A 1 R-induced IP accumulation

QL-induced IP accumulation

  

Basal

10 μM CHA

Fold Stimulation

Fold Stimulation

pcDNA1

N/A

10 ± 3

11 ± 2

1.1 ± 0.2

N/A

16

Yes

23 ± 5

237 ± 13

10.0 ± 0.6*

7.8 ± 0.4 #

N30

Yes

23 ± 4

81 ± 10

3.5 ± 0.7*

6.5 ± 0.4 #

N102

Yes

17 ± 4

13 ± 4

0.8 ± 0.2

1.1 ± 0.1

N155

Yes

13 ± 3

10 ± 1

0.8 ± 0.1

1.0 ± 0.1

N188

Yes

19 ± 4

155 ± 13

8.2 ± 0.3*

4.2 ± 0.2 #

N200

Yes

11 ± 3

15 ± 2

1.3 ± 0.1

2.0 ± 0.2 #

N210

Yes

12 ± 3

10 ± 2

0.8 ± 0.2

1.3 ± 0.1

N246

No

10 ± 2

10 ± 1

1.0 ± 0.1

0.8 ± 0.0

N266

No

27 ± 3

17 ± 2

0.6 ± 0.3

1.2 ± 0.2

N295

No

9 ± 1

9 ± 1

1.0 ± 0.1

0.9 ± 0.2

C25

Yes

15 ± 3

92 ± 7

6.1 ± 0.5*

4.1 a

C44

Yes

78 ± 11

183 ± 15

2.3 ± 0.2*

2.6 ± 0.1 #

C164

No

9 ± 2

8 ± 3

0.9 ± 0.3

1.3 ± 0.2

C174

No

13 ± 3

14 ± 9

1.1 ± 0.1

1.4 ± 0.2

C186

No

4 ± 1

6 ± 2

1.5 ± 0.5

1.4 ± 0.2

C219

No

7 ± 1

9 ± 2

1.3 ± 0.3

1.2 ± 0.1

C272

No

8 ± 1

9 ± 1

1.1 ± 0.1

1.2 ± 0.1

zβ2β3

Yes

21 ± 5

51 ± 2

3.0 ± 0.4*

4.3 ± 0.4 #

zβ3

Yes

14 ± 1

79 ± 5

5.6 ± 0.3*

6.9 ± 0.4 #

N200-C164

No

15 ± 1

19 ± 1

1.3 ± 0.1

0.9 ± 0.2

N188-C164

No

13 ± 0

14 ± 1

1.1 ± 0.0

1.0 ± 0.2

z

No

10 ± 1

9 ± 3

0.9 ± 0.3

1.1 ± 0.1

  1. HEK 293 cells were co-transfected with A1R and pcDNA1 or the indicated Gα subunit. Transfectants were labeled with [3H]inositol (2.5 μCi/mL) in DMEM containing 5% FBS overnight. IP3 formations were examined in the absence (basal) or presence of 10 μM CHA for 60 min. Fold stimulations were calculated as the ratios of CHA-induced to basal IP3 accumulations or QL-induced to wild-type IP3 accumulations. Data represent the mean ± S.D. of triplicate determinations of a single representative experiment, n = 3; significant responses are shown in bold and italic. Constructs containing an intact putative PLCβ binding domain (α2-β4-α3-β5 regions) of Gα16 are marked with a "Yes".
  2. * CHA-stimulated IP3 production is significantly greater than basal (DMSO vehicle); paired t-test, p ≤ 0.05.
  3. #QL-stimulated IP3 production is significantly greater than the corresponding wild-type activity; paired t-test, p ≤ 0.05.
  4. aQL-induced fold stimulation of 16z25 was extrapolated from [38].