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Chemical phosphorylation of myo-inositol
Sol/ Blol. Biochem. Vol. 16. No. 1. pp. 73-75. 1981
Printed in Gear
Britain. All rights reserved
CHEMICAL
Copyright
PHOSPHORYLATION JOHN EMSLEY
Department
of Chemistry.
King’s
OF MY0 -1NOSITOL
and SHAHIDA B. College,
Strand.
0038-0717 81 53.00+0.00 C 198-l Pergamon Press Ltd
NIAZI
London WC2R ZLS, U.K.
(Accepted 20 July 1982) Summary-Attempts to phosphorylate myo-inositol with various polyphosphates or urea-phosphate under aqueous conditions proved unsuccessful, despite reports to the contrary. The easiest way to phosphorylate myo-inositol completely is to heat it with H,PO, under reduced pressure at 1jO’C for 6 h.
INTRODUCTION
In citro phosphorylation
of alcohols and carbohydrates has been studied intensively for many years and generated a large body of literature (Cherbuliez, 1973; Emsley and Hall, 1976). One cyclitol, myoinositol, IH,, has proved particularly resistant to chemical phosphorylating reagents, although its natural abundance as the hexaphosphate, IP,, in plants and especially in soils is so great as to make it possibly the largest single component of pedosphere phosphorus (Anderson, 1967; Emsley, 1980). It has been reported that myo-IH, can be phosphorylated partly or completely by a variety of chemical reagents under aqueous and non-aqueous conditions but it is clear that these reports must be viewed with some scepticism (A. F. Harrison, personal communication). Researchers wishing to prepare “P-1abelled IP6 resort to the more reliable in ciao method using the mung bean method (Martin, 1970). We have examined the various methods suggested for the chemical phosphorylation of myo-IH, using principally the polyphosphates, which are thought to be involved in the metabolic phosphorylation process. MATERIALS
AND IMETHODS
Reaction of myo-inositol and phosphoric acid
myo-Inositol (4.00 g, 22 mmol) and H,PO, (14.8 g 85% solution; 132 mmol) were heated at 130X for 10 h under reduced pressure (water pump, ca. 2 kPa). The solution was cooled, diluted with water and barium acetate solution added which formed a precipitate that was redissolved in H,PO, at pH 2 and reprecipitated. The “P NMR spectrum of the redissolved material showed it to be mainly myo-inositol hexaphosphate by the 1:2:2: 1 collection of signals centered at 2.00 ppm (Emsley and Niazi, 1981) with’ small amounts ( c 5%) of inorganic monophosphate and diphosphate (6 - 9 ppm). The reprecipitated barium salt analysed as follows: found, C, 5.09; P, 12.63%. C6H,Ba,P,02, requires C, 4.90; P, 12.65%. “C NMR and TLC showed the presence of other myo-inositol phosphates in small amounts. When the same reaction is performed at 150°C there is almost quantitative conversion to IP6 ( > 95%) after 6 h. Reaction of myo-inositol and NH,[H2P04]
Reagents
myo-Inositol was 85% H,PO,, urea (NH,)JHPO,] and BDH; Na,P,O,, by
ical shifts of signals measured in ppm from 85% H,PO, as external reference standard. Other specifications were: pulse width 12.5~s; number of points 8 K; time constant - 1 to -2 sweep width 6000 Hz; offset 4500 Hz; flip angle 30’.
supplied by BDH Biochemicals; and NH,[H,PO,] by Fisons; sodium hexametaphosphate by Hopkin & Williams.
Chromatography
Thin layer chromatography (TLC) was done using Merck plastic sheets coated with microcrystalline cellulose F25a (0.1 mm) and with the solvent system I-propanol-NH,-HI0 (5:4: 1). Phosphate zones were detected with 60% HClO,-ammonium molybdateHCI-acetone solution and R, values compared with those of Cosgrove (1980; Chap. 3, pp. 12-25). “P NMR spectroscop)
“P NMR spectra were recorded on a Bruker HFX 90 spectrometer operating at 36.4 MHz at 307 + 2 K. Solutions were prepared with about 5% phosphate content to which a few drops of D,O were added as a lock. Broad band decoupling and pulsed Fourier transform (512-2048 scans) were used and the chem-
myo -1nositol (1 .OOg, 5.5 mmol) and NH,[H,PO,] (3.83 g, 33 mmol) were ground together and heated. The mixture melted at 18O’C, whereupon the temperature was lowered to 15O’C where it remained fluid and the heating continued under reduced pressure (water pump) for 10 h. The mixture was cooled, dissolved in water and phosphates precipitated by the addition of barium acetate solution. By techniques and analyses described in the previous reaction the product was shown to be IP, with some inorganic monophosphate and about 20% diphosphate. Reaction of myo-inositol and (NH,),
[HPO,]
myo-Inositol(1.00 g, 5.5 mmol) and (NH,)? [HPO,] (4.40 g, 33 mmol) were ground together and heated. Melting occurred, with evolution of NH!, at 125°C. The reaction mixture then solidified to a fused mass which was kept at 14O’C for 18 h, samples also being removed after 6 and 12 h. By techniques and analyses described in the first reaction it was seen that inorganic monophosphate had all been consumed by
JOHN EMSLEY and SHAHIDA B. NIAZI
Table I. The r:ac~on of mw-mositol
and inorganic phosphor?lating
reagents in aqueous solution’
Reagent
Conditions
Phosphorylation [analysis]
88”, H,PO,
150 C: 21 h
None [“P NMR]
No change
ReRur; 5 h: NH&I’
None [“P SMR]
Some diphosphate formed (5’J
Reflur: 20 h: (SH,),CO
None [“P NiMR]
pH 4.7 and IO: 15h: reflux”
None [“P NMR]
Some hydrolysis of triphosphate
(NaPOI),
pH 10: 15h reflux
None [“P NMR]
Some hydrolysis of polyphosphate
Na,P,O,’
pH I?; 4 months: room temperature
None [“P NMR]
No change
Na,P,O,’
pH IO: I months: room temperature
None [“P NMR]
Some hydrolysis to di- and monophosphate
Na,P,O,’
pH II; 15h; reflux
None [j’P NMR]
Some hydrolysis to di- and monophosphate
(NH:),CO.H,PO, NH,[H:PO,]
N.%P,O,,,
Remarks
Trace of diphosphats (<
I”,)
“IH, concentration I M. ratio IH,: phosphorylating agent = I :6. %rea phosphate. ‘Suggested catalyst for urea phosphate (Lohrman and Orgel. 1971) ‘?he addition of Mg” as a suggested catalyst has no effect. ‘Sodium “hexametaphosphate“ (so-called commercially). ‘Sodium cyclotriphosphate (also referred to as sodium trimetaphosphate).
18 h; diphosphate, which was present at 6 and 12 h, was also absent at the end. The product was IP+ Reaction of myo-inositol and phosphate etc, in water
Details of these reactions, none of which was successful in producing any inositol phosphate, are summarized in Table 1. RESULTS AND DISCUSSION
Undoubtedly the partial phosphorylation of IH6 can be achieved using POCI, (Kiely et a/., 1974) or N-benzoylphosphoramidic acid (Anderson et al., 1969; Angyal and Russell, 1969). Both reagents require non-aqueous conditions. The partial phosphorylation of IH6 in water has been claimed to occur using sodium cyclotriphosphate, Na,P,09 (sodium trimetaphosphate), when mono-, and di-phosphate of myo-inositol were identified after 40 h at room temperature (Cos rove, 1972). This reagent is capable of phosphory ! atmg 1967) and ribonucleosides alcohols (Feldman, (Safihill, 1970; Schwartz, 1969). Other polyphosphates would therefore seem to be serious contenders for aqueous phosphorylation, such as sodium triphosphate, Na,P,O,,, which has been used to phosphorylate adenosine (Schwartz and Ponnamperuma, 1968), and even monophosphates in the presence of urea at 65-IOO’C will phosphorylate uridine to a significant extent after 24 h (Lohrman and Orgel, 1971). Complete phosphorylation of IH6 by chemical means is a controversial subject. Posternak (1921) was the first to claim the synthesis of IP6 using polyphosphoric acid, PPA, (H3POI + P,O,,J and achieved a yield of ca. 8%. This work was challenged at the time (Anderson, 1920) and later (Fowler. 1956), the suggestion being that partial but not complete phosphorylation was possible by this method.
By using PPA in a sealed tube and heating to 15O’C Cosgrove (1975) reported the complete phosphorylation of epi- and nrltco-inositols, although heating for 24 h caused some epimerization. Under the experimental conditions reported here, no evidence has been obtained that IH, is phosphorylated in aqueous medium by any of the reagents shown in Table I. “‘P NMR spectroscopic analysis, the ideal investigative tool for such conditions (Newman and Tate, 1980), shows some diphosphate formation from NH,[HzPO,]-urea but no observable production of any inositol phosphates. It clearly revealed that under aqueous conditions the polyphosphates were hydrolysed to lower phosphates. Claims to phosphorylate inositol, even partially, under aqueous conditions must be treated with some scepticism. Inositol hexaphosphate can be produced by heating together 85% H;PO, and IH, at IX-15O’C for several hours-under reduced pressure. This condition removes water which leads to the formation of PPA (equation I) which may be the active phosphorylating agent. n H,PO,-*H,
+?P,,03, _ , + (n - 1)HzOt
(1) However equation (1) is a slow reaction and by itself H,PO, produces only 6:/, H,P,O? after 10 h rising to 44% after 25 h when ca. IO’?
(2)
It may also be possible under these conditions, since IP, is produced in > 907; yield after only 6 h, for H3P0, itself to phosphorylate partially phosphorylated inositol directly. This it could do by first condensing with a phosphate group already attached to the inositol ring, followed by attack at an adjacent
Phosphorylation
hydroxyl group: equation (3). where P* represents a phosphate group. IHSP* + H,PO+-IHjP*<*
+ H?Ot)
(3)
In this manner Cosgrove explained the epimerization of epi-inositol, and in the method proposed here continued heating for 21 h produced a product whose “P NMR spectrum was not the same as that of m_~o-IP,, again suggestive of epimerization. In addition to H,PO, the use of ammonium phosphates is capable of phosphorylating IH, to IP, at 15O’C. These reactions are assisted by the presence of urea which is capable of converting these salts to ammonium polyphosphates (Emsley and Niazi, 1982).
dcX_nonledgemenr-We thank the Government of Pakistan for a research grant for S.B.N.
of inositol
15
and the epimerization of m.vo-inositol pentaphosphates, Soil Biology & Biochemisrry 4. 387-396. Cosgrove D. J. (1975) The phosphorylation
and muco-inositol
with polyphosphotic
of epi-inositol acid. Carbo-
hydrare Research 40, 380-384.
Cosgrove D. J. (1980) lnositol Phosphares. Else\-ier. Amsterdam. Emsley J. (1980) The phosphorus cycle. In The Handbook of Encironmenfat Chemisrry. Vol. I Part A, (0. Hutzinger, Ed.) pp. 147-167. Springer-Verlag. Berlin. Emsley J. and Hall C. D. (1976) The Chemisrry of Phosphorus. pp. 338-345. Harper & Row. London. Emsley J. and Niazi S. B. (1981) The structure of mroinositol hexaphosphate in solution: “P n.m.r. finvestigation. Phosphorus and Sulfur 10, 401107. Emsley J. and Kiazi S. B. (1982) Condensation of ammonium phosphates with urea at 12O’C. Journal of the Chemical Society. Dalton Transactions. pp. 2527-253 I, Feldman W. (1967) Das Trimetaphosphat als Triphosphorvieruncsmittel fur Alkohole und Kohlenhvdrate in *,.-. wassnnger Ldsung. Seine Sonderstellung unter den kondensierten Phosphaten. Chemische Berichre 100, 3850-3860. Fowler H. D. (1956) Characterization of phytin in peas. Journal of the Science of Food and dgriculrure 7. 38 l-386.
REFERESCES Anderson G. (1967) Nucleic acids, derivatives and organic phosphates, Soil Biology & Biochemistry 1. 67-90. Anderson R. J. (1920) S$thesis of phytic acid. Journal of Biological Chemistry 43. 117-128. Anderson S. J., Luttrell B. M., Russell A. F. and Rutherford D. (1969) Inositol phosphates. glycosides. and some S-carbon cyclitols. Annals of [he New York Academy of Sciences 165, 533-540.
Angyal S. J. and Russell A. F. (1969) Cyclitols XXIX, Polyphosphorylation and polyols. The synthesis of Maoinositol pentaphosphates. Australian Journal ofChemiswy 22, 391-404.
Cherbuliez E. (1973) Organic derivatives (esters and organic anhydro acids) of phosphoric and polyphosphoric acids. In Organic Phosphorus Compounds (G. M. Kosolapoff and L. Maier, Eds). pp. 21 l-157. Wiley-Interscience, New York. Cosgrove D. J. (1972) The origin of inositol polyphosphates in soil. Some model experiments in aqueous systems involving the chemical phosphorylation of m.ro-inositol
Kiely D. E., Abruscato G. J. and Baburao V. (1974) A synthesis of (+) myo-inositol l-phosphate. Carhohydrafe Research 34, 307-3 13.
Lohrman R. and Orgel L. E. (1971) Urea-inorganic phosphate mixtures as prebiotic phosphorylating agents. Science 171, 490-494. Martin J. K. (1970) Preparation of ‘:P-labelled inositol hexaphosphate. Analytical Biochemirry 36, 233-237. Newman R. H. and Tate K. R. (1980) Soil characterization by “P nuclear magnetic resonance. Communications in Soil Science and Plant AnatFsis 11, 835-838.
Posternak S. (1921) Symhesis of inosite heaaphosphoric acid. Journal of Biological Chemisrry 46, 433-G7. SafIhill R. (1970) Selective phosphorylation of the cis-2’.3’with triribonucleosides diol of unprotected metaphosphate in aqueous solution. Journal of Organic Chemistry 35, 2881-2883.
Schwartz A. W. (1969) Specific phosphorylation of the 2’and 3’-oositions in ribonucleosides. Chemical Communicarion;, 1393.
Schwartz A. and Ponnamperuma C. (1968) Phosphorylation of adenosine with linear polyphosphate salts in aqueous solution. iVoture 218, 443.
Printed in Gear
Britain. All rights reserved
CHEMICAL
Copyright
PHOSPHORYLATION JOHN EMSLEY
Department
of Chemistry.
King’s
OF MY0 -1NOSITOL
and SHAHIDA B. College,
Strand.
0038-0717 81 53.00+0.00 C 198-l Pergamon Press Ltd
NIAZI
London WC2R ZLS, U.K.
(Accepted 20 July 1982) Summary-Attempts to phosphorylate myo-inositol with various polyphosphates or urea-phosphate under aqueous conditions proved unsuccessful, despite reports to the contrary. The easiest way to phosphorylate myo-inositol completely is to heat it with H,PO, under reduced pressure at 1jO’C for 6 h.
INTRODUCTION
In citro phosphorylation
of alcohols and carbohydrates has been studied intensively for many years and generated a large body of literature (Cherbuliez, 1973; Emsley and Hall, 1976). One cyclitol, myoinositol, IH,, has proved particularly resistant to chemical phosphorylating reagents, although its natural abundance as the hexaphosphate, IP,, in plants and especially in soils is so great as to make it possibly the largest single component of pedosphere phosphorus (Anderson, 1967; Emsley, 1980). It has been reported that myo-IH, can be phosphorylated partly or completely by a variety of chemical reagents under aqueous and non-aqueous conditions but it is clear that these reports must be viewed with some scepticism (A. F. Harrison, personal communication). Researchers wishing to prepare “P-1abelled IP6 resort to the more reliable in ciao method using the mung bean method (Martin, 1970). We have examined the various methods suggested for the chemical phosphorylation of myo-IH, using principally the polyphosphates, which are thought to be involved in the metabolic phosphorylation process. MATERIALS
AND IMETHODS
Reaction of myo-inositol and phosphoric acid
myo-Inositol (4.00 g, 22 mmol) and H,PO, (14.8 g 85% solution; 132 mmol) were heated at 130X for 10 h under reduced pressure (water pump, ca. 2 kPa). The solution was cooled, diluted with water and barium acetate solution added which formed a precipitate that was redissolved in H,PO, at pH 2 and reprecipitated. The “P NMR spectrum of the redissolved material showed it to be mainly myo-inositol hexaphosphate by the 1:2:2: 1 collection of signals centered at 2.00 ppm (Emsley and Niazi, 1981) with’ small amounts ( c 5%) of inorganic monophosphate and diphosphate (6 - 9 ppm). The reprecipitated barium salt analysed as follows: found, C, 5.09; P, 12.63%. C6H,Ba,P,02, requires C, 4.90; P, 12.65%. “C NMR and TLC showed the presence of other myo-inositol phosphates in small amounts. When the same reaction is performed at 150°C there is almost quantitative conversion to IP6 ( > 95%) after 6 h. Reaction of myo-inositol and NH,[H2P04]
Reagents
myo-Inositol was 85% H,PO,, urea (NH,)JHPO,] and BDH; Na,P,O,, by
ical shifts of signals measured in ppm from 85% H,PO, as external reference standard. Other specifications were: pulse width 12.5~s; number of points 8 K; time constant - 1 to -2 sweep width 6000 Hz; offset 4500 Hz; flip angle 30’.
supplied by BDH Biochemicals; and NH,[H,PO,] by Fisons; sodium hexametaphosphate by Hopkin & Williams.
Chromatography
Thin layer chromatography (TLC) was done using Merck plastic sheets coated with microcrystalline cellulose F25a (0.1 mm) and with the solvent system I-propanol-NH,-HI0 (5:4: 1). Phosphate zones were detected with 60% HClO,-ammonium molybdateHCI-acetone solution and R, values compared with those of Cosgrove (1980; Chap. 3, pp. 12-25). “P NMR spectroscop)
“P NMR spectra were recorded on a Bruker HFX 90 spectrometer operating at 36.4 MHz at 307 + 2 K. Solutions were prepared with about 5% phosphate content to which a few drops of D,O were added as a lock. Broad band decoupling and pulsed Fourier transform (512-2048 scans) were used and the chem-
myo -1nositol (1 .OOg, 5.5 mmol) and NH,[H,PO,] (3.83 g, 33 mmol) were ground together and heated. The mixture melted at 18O’C, whereupon the temperature was lowered to 15O’C where it remained fluid and the heating continued under reduced pressure (water pump) for 10 h. The mixture was cooled, dissolved in water and phosphates precipitated by the addition of barium acetate solution. By techniques and analyses described in the previous reaction the product was shown to be IP, with some inorganic monophosphate and about 20% diphosphate. Reaction of myo-inositol and (NH,),
[HPO,]
myo-Inositol(1.00 g, 5.5 mmol) and (NH,)? [HPO,] (4.40 g, 33 mmol) were ground together and heated. Melting occurred, with evolution of NH!, at 125°C. The reaction mixture then solidified to a fused mass which was kept at 14O’C for 18 h, samples also being removed after 6 and 12 h. By techniques and analyses described in the first reaction it was seen that inorganic monophosphate had all been consumed by
JOHN EMSLEY and SHAHIDA B. NIAZI
Table I. The r:ac~on of mw-mositol
and inorganic phosphor?lating
reagents in aqueous solution’
Reagent
Conditions
Phosphorylation [analysis]
88”, H,PO,
150 C: 21 h
None [“P NMR]
No change
ReRur; 5 h: NH&I’
None [“P SMR]
Some diphosphate formed (5’J
Reflur: 20 h: (SH,),CO
None [“P NiMR]
pH 4.7 and IO: 15h: reflux”
None [“P NMR]
Some hydrolysis of triphosphate
(NaPOI),
pH 10: 15h reflux
None [“P NMR]
Some hydrolysis of polyphosphate
Na,P,O,’
pH I?; 4 months: room temperature
None [“P NMR]
No change
Na,P,O,’
pH IO: I months: room temperature
None [“P NMR]
Some hydrolysis to di- and monophosphate
Na,P,O,’
pH II; 15h; reflux
None [j’P NMR]
Some hydrolysis to di- and monophosphate
(NH:),CO.H,PO, NH,[H:PO,]
N.%P,O,,,
Remarks
Trace of diphosphats (<
I”,)
“IH, concentration I M. ratio IH,: phosphorylating agent = I :6. %rea phosphate. ‘Suggested catalyst for urea phosphate (Lohrman and Orgel. 1971) ‘?he addition of Mg” as a suggested catalyst has no effect. ‘Sodium “hexametaphosphate“ (so-called commercially). ‘Sodium cyclotriphosphate (also referred to as sodium trimetaphosphate).
18 h; diphosphate, which was present at 6 and 12 h, was also absent at the end. The product was IP+ Reaction of myo-inositol and phosphate etc, in water
Details of these reactions, none of which was successful in producing any inositol phosphate, are summarized in Table 1. RESULTS AND DISCUSSION
Undoubtedly the partial phosphorylation of IH6 can be achieved using POCI, (Kiely et a/., 1974) or N-benzoylphosphoramidic acid (Anderson et al., 1969; Angyal and Russell, 1969). Both reagents require non-aqueous conditions. The partial phosphorylation of IH6 in water has been claimed to occur using sodium cyclotriphosphate, Na,P,09 (sodium trimetaphosphate), when mono-, and di-phosphate of myo-inositol were identified after 40 h at room temperature (Cos rove, 1972). This reagent is capable of phosphory ! atmg 1967) and ribonucleosides alcohols (Feldman, (Safihill, 1970; Schwartz, 1969). Other polyphosphates would therefore seem to be serious contenders for aqueous phosphorylation, such as sodium triphosphate, Na,P,O,,, which has been used to phosphorylate adenosine (Schwartz and Ponnamperuma, 1968), and even monophosphates in the presence of urea at 65-IOO’C will phosphorylate uridine to a significant extent after 24 h (Lohrman and Orgel, 1971). Complete phosphorylation of IH6 by chemical means is a controversial subject. Posternak (1921) was the first to claim the synthesis of IP6 using polyphosphoric acid, PPA, (H3POI + P,O,,J and achieved a yield of ca. 8%. This work was challenged at the time (Anderson, 1920) and later (Fowler. 1956), the suggestion being that partial but not complete phosphorylation was possible by this method.
By using PPA in a sealed tube and heating to 15O’C Cosgrove (1975) reported the complete phosphorylation of epi- and nrltco-inositols, although heating for 24 h caused some epimerization. Under the experimental conditions reported here, no evidence has been obtained that IH, is phosphorylated in aqueous medium by any of the reagents shown in Table I. “‘P NMR spectroscopic analysis, the ideal investigative tool for such conditions (Newman and Tate, 1980), shows some diphosphate formation from NH,[HzPO,]-urea but no observable production of any inositol phosphates. It clearly revealed that under aqueous conditions the polyphosphates were hydrolysed to lower phosphates. Claims to phosphorylate inositol, even partially, under aqueous conditions must be treated with some scepticism. Inositol hexaphosphate can be produced by heating together 85% H;PO, and IH, at IX-15O’C for several hours-under reduced pressure. This condition removes water which leads to the formation of PPA (equation I) which may be the active phosphorylating agent. n H,PO,-*H,
+?P,,03, _ , + (n - 1)HzOt
(1) However equation (1) is a slow reaction and by itself H,PO, produces only 6:/, H,P,O? after 10 h rising to 44% after 25 h when ca. IO’?
(2)
It may also be possible under these conditions, since IP, is produced in > 907; yield after only 6 h, for H3P0, itself to phosphorylate partially phosphorylated inositol directly. This it could do by first condensing with a phosphate group already attached to the inositol ring, followed by attack at an adjacent
Phosphorylation
hydroxyl group: equation (3). where P* represents a phosphate group. IHSP* + H,PO+-IHjP*<*
+ H?Ot)
(3)
In this manner Cosgrove explained the epimerization of epi-inositol, and in the method proposed here continued heating for 21 h produced a product whose “P NMR spectrum was not the same as that of m_~o-IP,, again suggestive of epimerization. In addition to H,PO, the use of ammonium phosphates is capable of phosphorylating IH, to IP, at 15O’C. These reactions are assisted by the presence of urea which is capable of converting these salts to ammonium polyphosphates (Emsley and Niazi, 1982).
dcX_nonledgemenr-We thank the Government of Pakistan for a research grant for S.B.N.
of inositol
15
and the epimerization of m.vo-inositol pentaphosphates, Soil Biology & Biochemisrry 4. 387-396. Cosgrove D. J. (1975) The phosphorylation
and muco-inositol
with polyphosphotic
of epi-inositol acid. Carbo-
hydrare Research 40, 380-384.
Cosgrove D. J. (1980) lnositol Phosphares. Else\-ier. Amsterdam. Emsley J. (1980) The phosphorus cycle. In The Handbook of Encironmenfat Chemisrry. Vol. I Part A, (0. Hutzinger, Ed.) pp. 147-167. Springer-Verlag. Berlin. Emsley J. and Hall C. D. (1976) The Chemisrry of Phosphorus. pp. 338-345. Harper & Row. London. Emsley J. and Niazi S. B. (1981) The structure of mroinositol hexaphosphate in solution: “P n.m.r. finvestigation. Phosphorus and Sulfur 10, 401107. Emsley J. and Kiazi S. B. (1982) Condensation of ammonium phosphates with urea at 12O’C. Journal of the Chemical Society. Dalton Transactions. pp. 2527-253 I, Feldman W. (1967) Das Trimetaphosphat als Triphosphorvieruncsmittel fur Alkohole und Kohlenhvdrate in *,.-. wassnnger Ldsung. Seine Sonderstellung unter den kondensierten Phosphaten. Chemische Berichre 100, 3850-3860. Fowler H. D. (1956) Characterization of phytin in peas. Journal of the Science of Food and dgriculrure 7. 38 l-386.
REFERESCES Anderson G. (1967) Nucleic acids, derivatives and organic phosphates, Soil Biology & Biochemistry 1. 67-90. Anderson R. J. (1920) S$thesis of phytic acid. Journal of Biological Chemistry 43. 117-128. Anderson S. J., Luttrell B. M., Russell A. F. and Rutherford D. (1969) Inositol phosphates. glycosides. and some S-carbon cyclitols. Annals of [he New York Academy of Sciences 165, 533-540.
Angyal S. J. and Russell A. F. (1969) Cyclitols XXIX, Polyphosphorylation and polyols. The synthesis of Maoinositol pentaphosphates. Australian Journal ofChemiswy 22, 391-404.
Cherbuliez E. (1973) Organic derivatives (esters and organic anhydro acids) of phosphoric and polyphosphoric acids. In Organic Phosphorus Compounds (G. M. Kosolapoff and L. Maier, Eds). pp. 21 l-157. Wiley-Interscience, New York. Cosgrove D. J. (1972) The origin of inositol polyphosphates in soil. Some model experiments in aqueous systems involving the chemical phosphorylation of m.ro-inositol
Kiely D. E., Abruscato G. J. and Baburao V. (1974) A synthesis of (+) myo-inositol l-phosphate. Carhohydrafe Research 34, 307-3 13.
Lohrman R. and Orgel L. E. (1971) Urea-inorganic phosphate mixtures as prebiotic phosphorylating agents. Science 171, 490-494. Martin J. K. (1970) Preparation of ‘:P-labelled inositol hexaphosphate. Analytical Biochemirry 36, 233-237. Newman R. H. and Tate K. R. (1980) Soil characterization by “P nuclear magnetic resonance. Communications in Soil Science and Plant AnatFsis 11, 835-838.
Posternak S. (1921) Symhesis of inosite heaaphosphoric acid. Journal of Biological Chemisrry 46, 433-G7. SafIhill R. (1970) Selective phosphorylation of the cis-2’.3’with triribonucleosides diol of unprotected metaphosphate in aqueous solution. Journal of Organic Chemistry 35, 2881-2883.
Schwartz A. W. (1969) Specific phosphorylation of the 2’and 3’-oositions in ribonucleosides. Chemical Communicarion;, 1393.
Schwartz A. and Ponnamperuma C. (1968) Phosphorylation of adenosine with linear polyphosphate salts in aqueous solution. iVoture 218, 443.