Investigation of water bound to photosystem I with multiquantum filtered 17 O nuclear magnetic resonance.
A new analytical approach was developed to characterize the properties of water molecules bound to macromolecules in solution using 17 O 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 H2 17 O 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 17 O 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 17 O 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.
Krzystyniak
M
M
Institute of Chemistry, Sekr. C 2, Technische Universität Berlin, Strasse des 17 Juni 135, D-10623 Berlin, Germany. krzystyniak@chem.tu-berlin.de
Shen
Gaozhong
G
Golbeck
John H
JH
Antonkine
Mikhail L
ML
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D000458:Cyanobacteria / Q000502:physiology; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010103:Oxygen Isotopes / Q000737:chemistry*; D045331:Photosystem I Protein Complex / Q000737:chemistry*; D011789:Quantum Theory; D014867:Water
D001426:Bacterial Proteins; D010103:Oxygen Isotopes; D045331:Photosystem I Protein Complex; D014867:Water