Page 44 - The Biochemistry of Inorganic Polyphosphates

WU095/Kulaev
WU095-02
Methods of polyphosphate assay
28 March 9, 2004 15:25 Char Count= 0

NDPα + NTPα
ex PP4
PP4
Pi
v
Pi NDPβ+NTPγ
PP3
NAD(H) PP3
Pi cyt
UDPG
UDPG
PEP
PM PP1
SP NTP
PP2

− − − − − −
Chemical shift, δ (ppm)
Figure 2.7 The 31 P NMR spectrum at 202.46 MHz of S. cerevisiae (adapted from Nicolay et al.,
1982;ShanksandBailey,1988;Beauvoitetal.,1991;Gonzalesetal.,2000):SP,sugarphosphate;Pi cyt ,
ex
v
cytoplasmic Pi; Pi , extracellular Pi; Pi , vacuolar Pi; PM, phosphomannan; NTPα and NDPα, α-
phosphate groups of nucleoside triphosphates and nucleoside diphosphates, respectively; NTPβ and
NDPβ, β-phosphate groups of nucleoside triphosphates and nucleoside diphosphates, respectively;
NTPγ, γ -phosphate group of nucleoside triphosphates; PP1, pyrophosphate and terminal phosphate
of PolyP; NAD(H), nicotinamide adenine dinucleotide; UDPG, uridinediphosphoglucose; PEP, phos-
phoenolpyruvate; PP2 and PP3, penultimate phosphates of PolyP; PP4, middle phosphates of PolyP.

chain length, [PP1] the terminal phosphate groups signal intensity and [PP4] the middle
phosphate groups signal intensity, respectively (Pilatus et al., 1989):

3 × [PP1] + [PP4]
n = 2 (2.1)
[PP1]
The reported data (Nicolay et al., 1982; Pilatus et al., 1989) show that the signal intensity
of the middle phosphate groups is proportional to the PolyP concentration. However, there is
no proportionality in the signals from PolyPs with different chain lengths: the contribution
of the middle phosphate groups to the total signal intensity in 31 P NMR spectra decreases
with the increasing length of the PolyP chain.
A special investigation was carried out to check the correlation between the decrease in
the 31 P NMR signal intensity of the middle phosphate groups and the chain length, and to
obtain an equation describing this dependence, thus making it possible to take this effect