PMID- 2159473
OWN - NLM
STAT- MEDLINE
DCOM- 19900613
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 265
IP  - 14
DP  - 1990 May 15
TI  - Purification and characterization of Go alpha and three types of Gi alpha after 
      expression in Escherichia coli.
PG  - 8243-51
AB  - Complementary DNAs for the G protein alpha subunits Gi alpha 1, Gi alpha 2, Gi 
      alpha 3, and Go alpha were expressed in Escherichia coli, and the four proteins 
      were purified to homogeneity. The recombinant proteins exchange and hydrolyze 
      guanine nucleotide, are ADP-ribosylated by pertussis toxin, and interact with 
      beta gamma subunits. The rates of dissociation of GDP from Gi alpha 1 and Gi 
      alpha 3 (0.03 min-1) are an order of magnitude slower than that from rGo alpha; 
      release of GDP from Gi alpha 2 is also relatively slow (0.07 min-1). However, the 
      values of kcat for the hydrolysis of GTP by rGo alpha and the three rGi alpha 
      proteins are approximately the same, about 2 min-1 at 20 degrees C. The 
      recombinant proteins restore inhibition of Ca2+ currents in pertussis 
      toxin-treated dorsal root ganglion neurons in response to neuropeptide Y and 
      bradykinin, indicating that the proteins can interact functionally with all 
      necessary components of at least one signal transduction system. The two 
      different receptors function with different arrays of G proteins to mediate their 
      responses, since all four G proteins restored responses to bradykinin, while Gi 
      alpha 2 was inactive with neuropeptide Y. Despite these results, high 
      concentrations of activated Gi alpha proteins are without effect on adenylyl 
      cyclase activity, either in the presence or absence of forskolin or Gs alpha, the 
      G protein that activates adenylyl cyclase. These results are consistent with the 
      hypothesis that G protein beta gamma subunits are primarily responsible for 
      inhibition of adenylyl cyclase activity.
FAU - Linder, M E
AU  - Linder ME
AD  - Department of Pharmacology, University of Texas Southwestern Medical Center, 
      Dallas 75235.
FAU - Ewald, D A
AU  - Ewald DA
FAU - Miller, R J
AU  - Miller RJ
FAU - Gilman, A G
AU  - Gilman AG
LA  - eng
GR  - DA02121/DA/NIDA NIH HHS/United States
GR  - DA02575/DA/NIDA NIH HHS/United States
GR  - GM34497/GM/NIGMS NIH HHS/United States
GR  - etc.
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  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Adenylate Cyclase Toxin)
RN  - 0 (Calcium Channels)
RN  - 0 (Neuropeptide Y)
RN  - 0 (Receptors, Neurotransmitter)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Virulence Factors, Bordetella)
RN  - 146-91-8 (Guanosine Diphosphate)
RN  - 20762-30-5 (Adenosine Diphosphate Ribose)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - 9007-49-2 (DNA)
RN  - EC 2.4.2.31 (Pertussis Toxin)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - EC 4.6.1.1 (Adenylyl Cyclases)
RN  - S8TIM42R2W (Bradykinin)
SB  - IM
MH  - Adenosine Diphosphate Ribose/metabolism
MH  - Adenylate Cyclase Toxin
MH  - Adenylyl Cyclases/metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Blotting, Western
MH  - Bradykinin/pharmacology
MH  - Calcium Channels/physiology
MH  - DNA/genetics
MH  - Escherichia coli/analysis/*genetics
MH  - GTP-Binding Proteins/*genetics/isolation & purification/metabolism
MH  - Ganglia, Spinal/drug effects/physiology
MH  - *Gene Expression
MH  - Guanosine Diphosphate/metabolism
MH  - Guanosine Triphosphate/metabolism
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Neuropeptide Y/pharmacology
MH  - Pertussis Toxin
MH  - Rats
MH  - Receptors, Neurotransmitter/physiology
MH  - Recombinant Proteins/isolation & purification
MH  - Signal Transduction
MH  - Virulence Factors, Bordetella/metabolism/pharmacology
EDAT- 1990/05/15 00:00
MHDA- 1990/05/15 00:01
CRDT- 1990/05/15 00:00
PHST- 1990/05/15 00:00 [pubmed]
PHST- 1990/05/15 00:01 [medline]
PHST- 1990/05/15 00:00 [entrez]
AID - S0021-9258(19)39064-7 [pii]
PST - ppublish
SO  - J Biol Chem. 1990 May 15;265(14):8243-51.