TY  - JOUR
AU  - Inoue, R.
AU  - Biehl, R.
AU  - Rosenkranz, T.
AU  - Fitter, J.
AU  - Monkenbusch, M.
AU  - Radulescu, A.
AU  - Farago, B.
AU  - Richter, D.
TI  - Large domain fluctuations on 50ns timescale enable catalytic activity in phoshpglycerate kinase
JO  - Biophysical journal
VL  - 99
IS  - 7
SN  - 0006-3495
CY  - New York, NY
PB  - Rockefeller Univ. Press
M1  - PreJuSER-16351
SP  - 2309 - 2317
PY  - 2010
N1  - T.R. acknowledges financial support by the International Helmholtz Research School on Biophysics and Soft Matter (BioSoft).
AB  - Large-scale domain motions of enzymes are often essential for their biological function. Phosphoglycerate kinase has a wide open domain structure with a hinge near the active center between the two domains. Applying neutron spin echo spectroscopy and small-angle neutron scattering we have investigated the internal domain dynamics. Structural analysis reveals that the holoprotein in solution seems to be more compact compared to the crystal structure but would not allow the functionally important phosphoryl transfer between the substrates if the protein were static. Brownian large-scale domain fluctuation dynamics on a timescale of 50 ns was revealed by neutron spin echo spectroscopy. The dynamics observed was compared to the displacement patterns of low-frequency normal modes. The displacements along the normal-mode coordinates describe our experimental results reasonably well. In particular, the domain movements facilitate a close encounter of the key residues in the active center to build the active configuration. The observed dynamics shows that the protein has the flexibility to allow fluctuations and displacements that seem to enable the function of the protein. Moreover, the presence of the substrates increases the rigidity, which is deduced from a faster dynamics with smaller amplitude.
KW  - Biocatalysis
KW  - Diffusion
KW  - Kinetics
KW  - Models, Molecular
KW  - Neutron Diffraction
KW  - Phosphoglycerate Kinase: chemistry
KW  - Phosphoglycerate Kinase: metabolism
KW  - Protein Structure, Secondary
KW  - Protein Structure, Tertiary
KW  - Saccharomyces cerevisiae: enzymology
KW  - Scattering, Small Angle
KW  - Structure-Activity Relationship
KW  - Time Factors
KW  - Phosphoglycerate Kinase (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:20923666
C2  - pmc:PMC3042550
UR  - <Go to ISI:>//WOS:000282850600037
DO  - DOI:10.1016/j.bpj.2010.08.017
UR  - https://juser.fz-juelich.de/record/16351
ER  -