Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer. / Jorgensen, Rasmus; Holliday, Nicholas D; Hansen, Jakob L; Vrecl, Milka; Heding, Anders; Schwartz, Thue W; Elling, Christian E.

In: Molecular Pharmacology, Vol. 73, No. 2, 2007, p. 349-58.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Jorgensen, R, Holliday, ND, Hansen, JL, Vrecl, M, Heding, A, Schwartz, TW & Elling, CE 2007, 'Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer', Molecular Pharmacology, vol. 73, no. 2, pp. 349-58. https://doi.org/10.1124/mol.107.038877

APA

Jorgensen, R., Holliday, N. D., Hansen, J. L., Vrecl, M., Heding, A., Schwartz, T. W., & Elling, C. E. (2007). Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer. Molecular Pharmacology, 73(2), 349-58. https://doi.org/10.1124/mol.107.038877

Vancouver

Jorgensen R, Holliday ND, Hansen JL, Vrecl M, Heding A, Schwartz TW et al. Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer. Molecular Pharmacology. 2007;73(2):349-58. https://doi.org/10.1124/mol.107.038877

Author

Jorgensen, Rasmus ; Holliday, Nicholas D ; Hansen, Jakob L ; Vrecl, Milka ; Heding, Anders ; Schwartz, Thue W ; Elling, Christian E. / Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer. In: Molecular Pharmacology. 2007 ; Vol. 73, No. 2. pp. 349-58.

Bibtex

@article{fd37ace0f90511ddb219000ea68e967b,
title = "Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer",
abstract = "To analyze the interaction between the neurokinin-1 (NK-1) receptor and G-protein coupled receptor kinases (GRKs), we performed bioluminescence resonance energy transfer(2) (BRET(2)) measurements between the family A NK-1 receptor and GRK2 and GRK5 as well as their respective kinase-inactive mutants. We observed agonist induced interaction of both GRK5 and GRK2 with the activated NK-1 receptor. In saturation experiments, we observed GRK5 to interact with the activated receptor in a monophasic manner while GRK2 interacted in a biphasic manner with the low affinity phase corresponding to receptor affinity for GRK5. Agonist induced GRK5 interaction with the receptor was dependent on intact kinase-activity, whereas the high affinity phase of GRK2 interaction was independent of kinase activity. We were surprised to find that the BRET(2) saturation experiments indicated that before receptor activation, the full-length NK-1 receptor, but not a functional C-terminal tail-truncated receptor, is preassociated with GRK5 in a relatively low-affinity state. We demonstrate that GRK5 can compete for agonist induced GRK2 interaction with the NK-1 receptor, whereas GRK2 does not compete for receptor interaction with GRK5. We suggest that GRK5 is preassociated with the NK-1 receptor and that GRK5, rather than GRK2, is a key player in competitive regulation of GRK subtype specific interaction with the NK-1 receptor.",
author = "Rasmus Jorgensen and Holliday, {Nicholas D} and Hansen, {Jakob L} and Milka Vrecl and Anders Heding and Schwartz, {Thue W} and Elling, {Christian E.}",
note = "Keywords: Amino Acid Sequence; Animals; Cell Line; Energy Transfer; G-Protein-Coupled Receptor Kinases; Humans; Luminescent Proteins; Molecular Sequence Data; Protein Binding; Receptors, Neurokinin-1; Renilla",
year = "2007",
doi = "10.1124/mol.107.038877",
language = "English",
volume = "73",
pages = "349--58",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

RIS

TY - JOUR

T1 - Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer

AU - Jorgensen, Rasmus

AU - Holliday, Nicholas D

AU - Hansen, Jakob L

AU - Vrecl, Milka

AU - Heding, Anders

AU - Schwartz, Thue W

AU - Elling, Christian E.

N1 - Keywords: Amino Acid Sequence; Animals; Cell Line; Energy Transfer; G-Protein-Coupled Receptor Kinases; Humans; Luminescent Proteins; Molecular Sequence Data; Protein Binding; Receptors, Neurokinin-1; Renilla

PY - 2007

Y1 - 2007

N2 - To analyze the interaction between the neurokinin-1 (NK-1) receptor and G-protein coupled receptor kinases (GRKs), we performed bioluminescence resonance energy transfer(2) (BRET(2)) measurements between the family A NK-1 receptor and GRK2 and GRK5 as well as their respective kinase-inactive mutants. We observed agonist induced interaction of both GRK5 and GRK2 with the activated NK-1 receptor. In saturation experiments, we observed GRK5 to interact with the activated receptor in a monophasic manner while GRK2 interacted in a biphasic manner with the low affinity phase corresponding to receptor affinity for GRK5. Agonist induced GRK5 interaction with the receptor was dependent on intact kinase-activity, whereas the high affinity phase of GRK2 interaction was independent of kinase activity. We were surprised to find that the BRET(2) saturation experiments indicated that before receptor activation, the full-length NK-1 receptor, but not a functional C-terminal tail-truncated receptor, is preassociated with GRK5 in a relatively low-affinity state. We demonstrate that GRK5 can compete for agonist induced GRK2 interaction with the NK-1 receptor, whereas GRK2 does not compete for receptor interaction with GRK5. We suggest that GRK5 is preassociated with the NK-1 receptor and that GRK5, rather than GRK2, is a key player in competitive regulation of GRK subtype specific interaction with the NK-1 receptor.

AB - To analyze the interaction between the neurokinin-1 (NK-1) receptor and G-protein coupled receptor kinases (GRKs), we performed bioluminescence resonance energy transfer(2) (BRET(2)) measurements between the family A NK-1 receptor and GRK2 and GRK5 as well as their respective kinase-inactive mutants. We observed agonist induced interaction of both GRK5 and GRK2 with the activated NK-1 receptor. In saturation experiments, we observed GRK5 to interact with the activated receptor in a monophasic manner while GRK2 interacted in a biphasic manner with the low affinity phase corresponding to receptor affinity for GRK5. Agonist induced GRK5 interaction with the receptor was dependent on intact kinase-activity, whereas the high affinity phase of GRK2 interaction was independent of kinase activity. We were surprised to find that the BRET(2) saturation experiments indicated that before receptor activation, the full-length NK-1 receptor, but not a functional C-terminal tail-truncated receptor, is preassociated with GRK5 in a relatively low-affinity state. We demonstrate that GRK5 can compete for agonist induced GRK2 interaction with the NK-1 receptor, whereas GRK2 does not compete for receptor interaction with GRK5. We suggest that GRK5 is preassociated with the NK-1 receptor and that GRK5, rather than GRK2, is a key player in competitive regulation of GRK subtype specific interaction with the NK-1 receptor.

U2 - 10.1124/mol.107.038877

DO - 10.1124/mol.107.038877

M3 - Journal article

C2 - 17986524

VL - 73

SP - 349

EP - 358

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 2

ER -

ID: 10485751