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Adenosine 5'-(beta, gamma-imino)triphosphate and guanosine 5'-O-(2-thiodiphosphate) do not necessarily provide non-phosphorylating conditions in adenylate cyclase studies
- Source :
- Biochemical and biophysical research communications. 126(3)
- Publication Year :
- 1985
-
Abstract
- Commercial preparations of adenosine 5′-(β, γ-imino)triphosphate (App(NH)p) were found to be contaminated with a GTP-like substance(s) as well as a phosphate donor(s) for GDP. Thus, when these preparations were used as substrate with no purification, GDP was as effective as GTP in promoting PGE 1 stimulation of human platelet adenylate cyclase. With purified App(NH)p as substrate, the effect of PGE 1 with GDP was reduced but still observable, while that with GTP was unaltered. PGE 1 also caused a stimulation in the presence of guanosine 5′-o-(2-thiodiphosphate) (GDPβS) with ATP as substrate. Both of the PGE 1 -stimulated activities observed with GDP and its analog were completely lost by the addition of UDP, thereby, inhibiting GTP formation catalized by membrane-associated nucleoside diphosphate kinase. The results demonstrate that the stimulatory effects of PGE 1 observed with GDP and App(NH)p, and with GDPβS and ATP were transphosphorylation dependent and, therefore, the analogs must be used with special caution in adenylate cyclase studies.
- Subjects :
- Blood Platelets
GTP'
Adenylyl Imidodiphosphate
Biophysics
Guanosine
Adenylate kinase
Stimulation
Biochemistry
Cyclase
Guanosine Diphosphate
Substrate Specificity
chemistry.chemical_compound
Adenosine Triphosphate
medicine
Humans
Alprostadil
Phosphorylation
Molecular Biology
Prostaglandins E
Cell Biology
Thionucleotides
Adenosine
Nucleoside-diphosphate kinase
Guanine Nucleotides
Enzyme Activation
chemistry
Guanosine Triphosphate
medicine.drug
Adenylyl Cyclases
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 126
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Biochemical and biophysical research communications
- Accession number :
- edsair.doi.dedup.....89e4d55565e41e1bfb2737378772322f