A new analytical approach was developed to characterize the properties of water molecules bound to macromolecules in solution using O17 nuclear magnetic resonance (NMR) relaxation. A combination of conventional (single-quantum) and triple-quantum filtered Hahn echo and inversion recovery measurements was employed. From measured relaxation rate constants, the fraction and the correlation time of bound H2O17 molecules and the relaxation rate constant of bulk water in solution were calculated. This was done by solving analytically a set of nonlinear equations describing the overall relaxation rate constants in the presence of chemical exchange between bulk and bound water. The analytical approach shows the uniqueness of the solution for a given set of three relaxation rate constants. This important result sheds light on the data reduction problem from O17 NMR experiments on biological systems. Water bound in photosystem I isolated from the wild type and rubA variant of the cyanobacterium Synechocystis species PCC 7002 was investigated for the first time. The analysis revealed that photosystem I isolated from the wild type binds 1720±110 water molecules, whereas photosystem I isolated from the rubA variant binds only 1310±170. The accuracy of the method proposed can be increased by further O17 enrichment. The methodology, established for the first time in this work, allows the study of a diverse range of biological samples regardless of their size and molecular weight. Applied initially to photosystem I, this novel method has important consequences for the future investigation of the assembly of biological molecules.

1.
K. L.
Britton
,
P. J.
Baker
,
M.
Fisher
,
S.
Ruzheinikov
,
D. J.
Gilmour
,
M.-J.
Bonete
,
J.
Ferrer
,
C.
Pire
,
J.
Esclapez
, and
D. W.
Rice
,
Proc. Natl. Acad. Sci. U.S.A.
103
,
4846
(
2006
).
2.
U. D.
Ramirez
and
D. M.
Freymann
,
Acta Crystallogr., Sect. D: Biol. Crystallogr.
62
,
15201534
(
2006
).
3.
B.
Bagchi
,
Chem. Rev. (Washington, D.C.)
105
,
3197
(
2005
).
4.
S.
Wiesner
,
E.
Kurian
,
F. G.
Prendrgast
, and
B.
Halle
,
J. Mol. Biol.
286
,
233
(
1999
).
5.
M.
Gottschalk
,
N. A.
Dencher
, and
B.
Halle
,
J. Mol. Biol.
311
,
605
(
2001
).
6.
V. P.
Denisov
,
P.
Jonsson
, and
B.
Halle
,
Nat. Struct. Mol. Biol.
6
,
253
(
1999
).
7.
K.
Modig
,
E.
Kurian
,
F.
Prendergast
, and
B.
Halle
,
Protein Sci.
12
,
2768
(
2003
).
8.
K.
Modig
,
M.
Rademacher
,
C.
Lucke
, and
B.
Halle
,
J. Mol. Biol.
332
,
965
(
2003
).
9.
G.
Otting
,
E.
Liepinsh
, and
K.
Wüthrich
,
Science
254
,
974
(
1991
).
10.
E.
Baguet
,
B.
Chapman
,
A. M.
Torres
, and
P. W.
Kuchel
,
J. Magn. Reson., Ser. B
111
,
1
(
1996
).
11.
A.
Lehoux
,
M.
Krzystyniak
, and
E.
Baguet
,
J. Magn. Reson.
148
,
11
(
2001
).
12.
C. W.
Chung
and
S.
Wimperis
,
Mol. Phys.
76
,
47
(
1992
).
13.
E.
Baguet
,
N.
Hennenbert
,
B. E.
Chapman
,
A. M.
Torres
, and
P. W.
Kuchel
,
Magn. Reson. Chem.
35
,
S47
(
1997
).
14.
E.
Baguet
and
N.
Hennenbert
,
Biophys. Chem.
77
,
111
(
1999
).
15.
A. M.
Torres
,
S. M.
Grieve
,
B. E.
Chapman
, and
P. W.
Kuchel
,
Biophys. Chem.
67
,
187
(
1997
).
16.
A. M.
Torres
,
S. M.
Grieve
, and
P. W.
Kuchel
,
Biophys. Chem.
70
,
231
(
1998
).
17.
S. M.
Grieve
,
B.
Wickstead
,
A. M.
Torres
,
P.
Styles
,
S.
Wimperis
, and
P. W.
Kuchel
,
Biophys. Chem.
73
,
137
(
1998
).
18.
P.
Canfield
,
M. G.
Dahlbom
,
N.
Hush
, and
J. R.
Reimers
,
J. Chem. Phys.
124
,
024301
(
2006
).
19.
R. E.
Blankenship
,
Molecular Mechanism of Photosynthesis
(
Blackwell Science
,
Oxford
,
2002
).
20.
P.
Jordan
,
P.
Fromme
,
H. T.
Witt
,
O.
Klukas
,
W.
Saenger
, and
N.
Krauß
,
Nature (London)
411
,
909
(
2001
).
21.
M. L.
Antonkine
and
J. H.
Golbeck
, in
Photosytem I: The Light-Driven Plastocyanin: Ferredoxin Oxidoreductose
(
Springer
,
New York
,
2006
), Chap. 8, pp.
79
98
.
22.
G.
Shen
,
J.
Zhao
,
S. K.
Reimer
,
M. L.
Antonkine
,
Q.
Cai
,
S. M.
Weiland
,
J.
Golbeck
, and
D. A.
Bryant
,
J. Biol. Chem.
277
,
20343
(
2002
).
23.
Y.
Pushkar
,
Y.
Karyagina
,
D.
Stehlik
,
S.
Brown
, and
A.
van der Est
,
J. Biol. Chem.
280
,
12382
(
2005
).
24.
Y.
Sakuragi
,
B.
Zybailov
,
G.
Shen
,
D.
Bryant
,
J. H.
Golbeck
,
B. A.
Diner
,
I.
Karygina
,
Y.
Pushkar
, and
D.
Stehlik
,
J. Biol. Chem.
280
,
12371
(
2005
).
25.
G.
Shen
,
M. L.
Antonkine
,
A.
van der Est
,
I. R.
Vassiliev
,
K.
Brettel
,
R.
Bittl
,
S. G.
Zech
,
J.
Zhao
,
D.
Stehlik
,
D. A.
Bryant
, and
J. H.
Goldbeck
,
J. Biol. Chem.
277
,
20355
(
2002
).
26.
P.
Voigt
and
E.-W.
Knapp
,
J. Biol. Chem.
278
,
51993
(
2003
).
27.
H.
Ishikita
and
E.-W.
Knapp
,
J. Biol. Chem.
278
,
52002
(
2003
).
28.
D.
Mauzerall
, in
Photosytem I: The Light-Driven Plastocyanin: Ferredoxin Oxidoreductose
(
Springer
,
New York
,
2006
), Chap. 33, pp.
571
580
.
29.
N. G.
Bukhov
,
S.
Govindachary
,
E. A.
Egorova
, and
R.
Carpentier
,
Planta
219
,
110
(
2004
).
30.
M. L.
Antonkine
,
P.
Jordan
,
P.
Fromme
,
N.
Krauß
,
J.
Golbeck
, and
D.
Stehlik
,
J. Mol. Biol.
327
,
671697
(
2003
).
31.
V. P.
Denisov
and
B.
Halle
,
Faraday Discuss.
103
,
227
(
1996
).
32.
B.
Halle
,
V. P.
Denisov
, and
K.
Venu
, in
Structure Computation and Dynamics in Protein NMR
,
Biological Magnetic Resonance
Vol.
17
(
Kluwer Academic/Plenum
,
New York
,
1999
).
33.
B.
Halle
and
H.
Wennerström
,
J. Magn. Reson.
44
,
89
(
1981
).
34.
L.
Picullel
and
B.
Halle
,
J. Chem. Soc., Faraday Trans.
82
,
401
(
1986
).
35.
B.
Halle
,
T.
Andersson
,
S.
Forsen
, and
B.
Lindman
,
J. Am. Chem. Soc.
103
,
500
(
1981
).
36.
T. E.
Bull
,
S.
Forsen
, and
D. L.
Turner
,
J. Chem. Phys.
70
,
3106
(
1979
).
37.
V. P.
Denisov
and
B.
Halle
,
J. Mol. Biol.
245
,
682
(
1995
).
38.
T.
Hiyama
and
B.
Ke
,
Biochim. Biophys. Acta
267
,
160
(
1972
).
39.
H.
van Gorkom
,
J.
Tamminga
, and
J.
Haveman
,
Biochim. Biophys. Acta
347
,
417
(
1974
).
40.
F.
Mueh
and
A.
Zouni
,
Biochim. Biophys. Acta
1708
,
219
(
2005
).
41.
K.
Modig
,
E.
Liepinsh
,
G.
Otting
, and
B.
Halle
,
J. Am. Chem. Soc.
126
,
102
(
2004
).
42.
I.
Bertini
and
C.
Luchinat
,
Coordination Chemistry Reviews
(
Elsevier
,
New York
,
1996
), Vol.
150
, Chap. 3, pp.
77
111
.
You do not currently have access to this content.