PMID- 10029519
OWN - NLM
STAT- MEDLINE
DCOM- 19990316
LR  - 20101118
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 8
DP  - 1999 Feb 23
TI  - Comparative analysis of the efficacy of A1 adenosine receptor activation of Gi/o 
      alpha G proteins following coexpression of receptor and G protein and expression 
      of A1 adenosine receptor-Gi/o alpha fusion proteins.
PG  - 2272-8
AB  - HEK293T cells were transiently transfected to express either the human A1
      adenosine receptor together with pertussis toxin-resistant cysteine-to-glycine
      forms of the alpha subunits of Gi1 (C351G), Gi2 (C352G), and Gi3 (C351G) and
      wild-type Go1alpha or fusion proteins comprising the A1 adenosine receptor and
      these Gi/o G proteins to compare A1 adenosine receptor agonist-mediated
      activation of these Gi family G proteins upon coexpression of individual Gi/o G
      proteins and receptor versus expression as receptor-G protein fusion proteins.
      Addition of the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) 
      to membranes of pertussis toxin-treated cells resulted in a
      concentration-dependent stimulation of [35S]GTPgammaS binding with comparable
      amounts of NECA required to produce half-maximal stimulation following
      transfection of A1 adenosine receptor and Gi/o G proteins either as fusion
      proteins or as separate polypeptides. However, the magnitude of agonist-mediated 
      activation of GTPgammaS binding was greatly enhanced by expressing the A1
      adenosine receptor and Gi family G proteins from chimaeric open reading frames.
      This observation was consistent following the study of more than 40 agonists. No 
      preferential activation of any G protein was observed with more than 40 A1
      receptor agonists following cotransfection of receptor with G protein or
      transfection of receptor-G protein fusion proteins. These studies demonstrate the
      utility of using fusion proteins to study receptor-G protein interaction, show
      that the A1 adenosine receptor couples equally well to the Gi/o G proteins
      Gi1alpha, G i2alpha, Gi3alpha, and Go1alpha, and demonstrate that for a range of 
      agonists there is no selectivity for activation of any particular A1 adenosine
      receptor-Gi/o G protein combination.
FAU - Wise, A
AU  - Wise A
AD  - Receptor Systems Unit, Medicines Research Centre, Glaxo Wellcome Research and
      Development, Stevenage, England, U.K.
FAU - Sheehan, M
AU  - Sheehan M
FAU - Rees, S
AU  - Rees S
FAU - Lee, M
AU  - Lee M
FAU - Milligan, G
AU  - Milligan G
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Purinergic P1 Receptor Agonists)
RN  - 0 (Receptors, Purinergic P1)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Virulence Factors, Bordetella)
RN  - 35920-39-9 (Adenosine-5'-(N-ethylcarboxamide))
RN  - 37589-80-3 (Guanosine 5'-O-(3-Thiotriphosphate))
RN  - EC 2.4.2.31 (Pertussis Toxin)
RN  - EC 3.6.5.1 (GTP-Binding Protein alpha Subunits, Gi-Go)
SB  - IM
MH  - Adenosine-5'-(N-ethylcarboxamide)/pharmacology
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites/drug effects
MH  - Cell Line
MH  - Embryo, Mammalian
MH  - GTP-Binding Protein alpha Subunits, Gi-Go/biosynthesis/genetics/*metabolism
MH  - Guanosine 5'-O-(3-Thiotriphosphate)/metabolism
MH  - Humans
MH  - Kidney
MH  - Molecular Sequence Data
MH  - Pertussis Toxin
MH  - Purinergic P1 Receptor Agonists
MH  - Rats
MH  - Receptors, Purinergic P1/biosynthesis/genetics/*physiology
MH  - Recombinant Fusion Proteins/*biosynthesis/chemical synthesis
MH  - Transfection
MH  - Virulence Factors, Bordetella/pharmacology
EDAT- 1999/02/25 00:00
MHDA- 1999/02/25 00:01
CRDT- 1999/02/25 00:00
PHST- 1999/02/25 00:00 [pubmed]
PHST- 1999/02/25 00:01 [medline]
PHST- 1999/02/25 00:00 [entrez]
AID - 10.1021/bi982054f [doi]
AID - bi982054f [pii]
PST - ppublish
SO  - Biochemistry. 1999 Feb 23;38(8):2272-8. doi: 10.1021/bi982054f.