Adenosine 5'-phosphorothioate. A nucleotide analog that is a substrate, competitive inhibitor, or regulator of some enzymes that interact with adenosine 5'-phosphate

Abstract

Adenosine 5 '-phosphorothioate, an analog of adenosine 5 '-phosphate, has been prepared from adenosine and thiophosphoryl chloride in triethyl phosphate in 65 % yield and isolated as the sodium salt. The adenosine 5 '-phosphate content of this product (measured with adenylate kinase) was less than 0.5% and the content of adenosine derivatives other than the 5'phosphorothioate (measured as unchanged material after treatment with adenylate deaminase or with 5'nucleotidase) was 1-3 %. Adenosine 5 '-phosphorothioate (0.34 mM) was phosphorylated by adenosine triphosphate in the presence of muscle adenylate kinase at less than 0.3% of the rate with adenosine 5'-phosphate at the same concentration, but 0.25 mM adenosine 5 '-phosphorothioate increased the adenosine 5 '-phosphate concentration for half-maximal activity, [SI, 5 , from 0.15 to 0.18 mM and decreased the maximal velocity (V, micromoles per minute per milligram of protein) from 120 to 91 without changing the slope (1.35) of the Hill plot. Adenosine 5 '-phosphorothioate was deaminated by adenylate deaminase from rat skeletal muscle. The Michaelis constant, K,, was 1.6 mM and V was 125, 9% of the maximal velocity with adenosine 5'-phosphate(V = 1330; K, = 0.9 mM). With 5'-nucleotidase from Crotalus venom liberation of adenosine from adenosine 5 '-phosphorothioate was much slower than that from adenosine 5 '-phosphate (V = 0.026 and 1.39, respectively) but K, for adenosine 5'-phosphorothioate N ucleoside 5 '-monophosphates and nucleoside 5 '-triphosphates, particularly AMP and ATP, are some of the most important regulators of metabolic processes (for reviews, see Krebs (1964) and Atkinson (1966)).

The preparation of thiophosphate analogs of GDP and their interaction with EF‐Tu

“…GMP (S) was prepared by the methods described for the preparation of AMP (S) [4], the mixed diastereomer of GDP(t~-S) as described for ADP(a- S) [1], and GDP(fl-S) as described for ADP(fl-S) [5]. Myosin subfragment-t was prepared by the method in [6], actin by the method in [7], and EF-Tu • GDP from E. coli (MRE 600) by the method currently used in this laboratory (Leberman et al, in preparation).…”

31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate.

“…Glycogen was freed from AMP as described previously (13 resonances of free and bound AMP are partially resolved ( Interaction of Phosphorylase b with the Allosteric Effector AMP-S. The difficulty in resolving AMP and pyridoxal-P resonances was overcome by using AMP-S, an even better activator (17).…”

The mononucleotide-dependent, nonantisense mechanism of action of phosphodiester and phosphorothioate oligonucleotides depends upon the activity of an ecto-5′-nucleotidase

“…The dNMPSs were synthesized by means of the modified method of Murray and Atkinson 13 and purified by high-performance liquid chromatography (HPLC). Purine bases (guanine, xanthine, hypoxanthine, and adenine), deoxyguanosine, inosine, and dNMPs were obtained from Sigma (St Louis, MO).…”