PMID- 6138091
OWN - NLM
STAT- MEDLINE
DCOM- 19831220
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 22
IP  - 19
DP  - 1983 Sep 13
TI  - Reconstitution of catecholamine-stimulated guanosinetriphosphatase activity.
PG  - 4357-62
AB  - beta-Adrenergic receptors were partially purified from turkey erythrocyte 
      membranes by alprenolol-agarose chromatography to 0.25-2 nmol/mg of protein, and 
      the stimulatory guanosine 5'-triphosphate (GTP) binding protein of adenylate 
      cyclase (Gs) was purified from rabbit liver. These proteins were reconstituted 
      into phospholipid vesicles by addition of phospholipids and removal of detergent 
      by gel filtration. This preparation hydrolyzes GTP to guanosine 5'-diphosphate 
      (GDP) plus inorganic phosphate (Pi) in response to beta-adrenergic agonists. The 
      initial rate of isoproterenol-stimulated hydrolysis is approximately 1 mol of GTP 
      hydrolyzed min-1 X mol-1 of Gs. This low rate may be limited by the 
      hormone-stimulated binding of substrate, since it is roughly equal to the rate of 
      binding of the GTP analogue guanosine 5'-O-(3-[35S] thiotriphosphate) [( 35S]GTP 
      gamma S) to Gs in the vesicles. Activity in the absence of agonist, or in the 
      presence of agonist plus a beta-adrenergic antagonist, is 8-25% of the 
      hormone-stimulated activity. Guanosinetriphosphatase (GTPase) is not saturated at 
      10 microM GTP, and the response to GTP is formally consistent either with the 
      existence of multiple Km's or of a separate stimulatory site for GTP. The GTPase 
      activity of Gs in vesicles is also stimulated by 50 mM MgCl2 in the presence or 
      absence of receptor. Significant GTPase activity is not observed with 
      Lubrol-solubilized Gs, although [35S]-GTP gamma S binding is increased by Lubrol 
      solubilization.
FAU - Brandt, D R
AU  - Brandt DR
FAU - Asano, T
AU  - Asano T
FAU - Pedersen, S E
AU  - Pedersen SE
FAU - Ross, E M
AU  - Ross EM
LA  - eng
GR  - F32 GM09363/GM/NIGMS NIH HHS/United States
GR  - R01 GM30355/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Receptors, Adrenergic, beta)
RN  - 0 (Receptors, Cell Surface)
RN  - EC 3.1.3.2 (Phosphoric Monoester Hydrolases)
RN  - EC 3.6.1.- (GTP Phosphohydrolases)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - EC 4.6.1.1 (Adenylyl Cyclases)
RN  - L628TT009W (Isoproterenol)
SB  - IM
MH  - Adenylyl Cyclases/*metabolism
MH  - Animals
MH  - Erythrocyte Membrane/*metabolism
MH  - GTP Phosphohydrolases/*blood
MH  - GTP-Binding Proteins
MH  - Isoproterenol/*pharmacology
MH  - Kinetics
MH  - Liver/enzymology
MH  - Phosphoric Monoester Hydrolases/*blood
MH  - Rabbits
MH  - Receptors, Adrenergic, beta/isolation & purification/*metabolism
MH  - Receptors, Cell Surface/isolation & purification/*metabolism
MH  - Turkeys
EDAT- 1983/09/13 00:00
MHDA- 1983/09/13 00:01
CRDT- 1983/09/13 00:00
PHST- 1983/09/13 00:00 [pubmed]
PHST- 1983/09/13 00:01 [medline]
PHST- 1983/09/13 00:00 [entrez]
AID - 10.1021/bi00288a002 [doi]
PST - ppublish
SO  - Biochemistry. 1983 Sep 13;22(19):4357-62. doi: 10.1021/bi00288a002.