The stimulatory effect of vasoactive intestinal peptide (VIP) on the intracellular calcium concentration ([Ca2+]i) has been investigated in Chinese hamster ovary cells stably transfected with the reporter gene aequorin, and expressing human VPAC1, VPAC2, chimaeric VPAC1/VPAC2 or mutated receptors. The VIP-induced increase in [Ca2+]i was linearly correlated with receptor density, and was higher in cells expressing VPAC1 receptors than in cells expressing a similar density of VPAC2 receptors. The study was performed to establish the receptor sequence responsible for this difference. VPAC1/VPAC2 chimaeric receptors were first used for broad positioning: those receptors having the third intracellular loop (IC3) of the VPAC1 or the VPAC2 receptor behaved, in this respect, phenotypically like VPAC1 and VPAC2 receptors respectively. Replacement in the VPAC2 receptor of the sequence comprising residues 315–318 (VGGN) within IC3 by its VPAC1 receptor counterpart (residues 328–331; IRKS) and the introduction of VGGN instead of IRKS into VPAC1 was sufficient to mimic VPAC1 and VPAC2 receptor characteristics respectively. Thus a small sequence in the IC3 domain of the VPAC1 receptor is responsible for the efficient agonist-stimulated increase in [Ca2+]i.
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Conference Article|
August 01 2002
A small sequence in the third intracellular loop of the VPAC1 receptor is responsible for its efficient coupling to the calcium effector
I. Langer;
I. Langer
1
1Department of Biological Chemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat GE, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium
1To whom correspondence should be addressed (e-mail ilanger@ulb.ac.be)
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P. Vertongen;
P. Vertongen
1Department of Biological Chemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat GE, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium
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J. Perret;
J. Perret
1Department of Biological Chemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat GE, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium
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M. Waelbroeck;
M. Waelbroeck
1Department of Biological Chemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat GE, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium
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P. Robberecht
P. Robberecht
1Department of Biological Chemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat GE, CP 611, 808 Route de Lennik, B-1070 Brussels, Belgium
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Publisher: Portland Press Ltd
Received:
March 08 2002
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2002 Biochemical Society
2002
Biochem Soc Trans (2002) 30 (4): 447–450.
Article history
Received:
March 08 2002
Citation
I. Langer, P. Vertongen, J. Perret, M. Waelbroeck, P. Robberecht; A small sequence in the third intracellular loop of the VPAC1 receptor is responsible for its efficient coupling to the calcium effector. Biochem Soc Trans 1 August 2002; 30 (4): 447–450. doi: https://doi.org/10.1042/bst0300447
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