Studies of Phosphorylation. IV. The Phosphorylation of Nucleosides with Phosphorus Trihalide

Abstract

The treatment of a 2′,3′-O-isopropylidene nucleoside with phosphorus trichloride in the presence of trimethyl phosphate and subsequent hydrolysis gave a nucleoside 5′-phosphite in an excellent yield. Triaryl phosphates, trialkylphosphine oxides, and trialkyl phosphites were also found to be potential accelerators of the reaction. Their effects were comparable to that of trimethyl phosphate. The similar treatment of a 5′-O-acetyl nucleoside gave mixed nucleoside 2′- and 3′-phosphites in an approximate mole ratio of 4 : 6. The reaction intermediate, nucleoside phosphorodichloridite, was subjected to partial hydrolysis, giving the corresponding monochloridite. The product was readily oxidized to give the 5′-nucleotide. Chlorine was generally suitable as an oxidizing agent, but iodine gave better results for guanosine derivatives than did chlorine or bromine. The treatment of an unprotected nucleoside with phosphorus oxychloride in a trialkyl phosphate, prior to the reaction with phosphorus trichloride, could be used for the synthesis of a nucleoside 2′(or 3′),5′-diphosphate in a good yield.

The Synthesis of Three AMP‐Analogues: N6‐(6‐Aminohexyl)‐adenosine 5′‐Monophosphate, N6‐(6‐Aminohexyl)‐adenosine 2′,5′‐Bisphosphate, and N6‐(6‐Aminohexyl)‐adenosine 3′,5′‐Bisphosphate and Their Application as General Ligands in Biospecific Affinity Chromatography

“…P = phosphate that described for the preparation of N6-(6-aminohexy1)-adenosine 5'-monophosphate [6] except that a different phosphorylation method was used. This method, which is based on a recent report on the phosphorylation of several common nucleotides [14] yields a mixture of the isomers 6-chloropurine-riboside 2',5'-bisphosphate and 6-chloropurine-riboside 3',5'-bisphosphate. This mixture is difficult to resolve quantitatively by ion-exchange chromatography unless conversion to the corresponding aminohexyl derivatives is effected.…”

Addition of mononucleotides to oligoribonucleotide acceptors with T4 RNA ligase.

“…Adenosine 5',3'-diphosphate and adenosine 5',2'-diphosphate were from Sigma Chemical Co. and thymidine 5',3'-diphosphate was from Worthington Biochemical Co. Cytidine 5',3'(or 2')-diphosphate, guanosine 5',3'(2')-diphosphate, and uridine 5',3'-(2')-diphosphate were synthesized chemically by the method of Yoshikawa et al (13). Snake venom phosphodiesterase was purchased from Worthington Biochemical Co. Nuclease P1 and poly(C) were from Yamasa Shoyu Co. RNase T2 was obtained from Sankyo Co. and nuclease SW (silkworm nuclease) was from Seikagaku Kogyo Co. Bacterial alkaline phosphatase was prepared from Escherichia coli A19 cells by the method of Garen and Levinthal (14).…”