TY - JOUR
T1 - The use of the time-resolved X-ray solution scattering for studies of globular proteins
AU - Kuwajima, Kunihiro
AU - Arai, Munehito
AU - Inobe, Tomonao
AU - Ito, Kazuki
AU - Nakao, Masaharu
AU - Maki, Kosuke
AU - Kamagata, Kiyoto
AU - Kihara, Hiroshi
AU - Amemiya, Yoshiyuki
PY - 2002
Y1 - 2002
N2 - In order to improve the low signal-to-noise ratio of the time-resolved small-angle X-ray scattering, we have used a two-dimensional X-ray detector with a beryllium-windowed X-ray image intensifier and a charge-coupled device as an image sensor, and applied this to studies on (1) the kinetic folding reaction of α-lactalbumin, which accumulates the molten globule-like intermediate at an early stage of refolding and (2) the cooperative conformational transition of Escherichia coli chaperonin GroEL induced by ATP, which occurs in an allosteric manner between the close and open conformational states. In the α-lactalbumin reaction, we have firmly established the equivalence between the kinetic intermediate and the equilibrium molten globule state, and obtained further information about dehydration from the highly hydrated folding intermediate during a late stage of refolding. In the chaperonin study, we have successfully observed the kinetics of the allosteric transition of GroEL that occurs with a rate constant of about 3-4 s-1 at 5°C. The combination of the time-resolved X-ray scattering with other spectroscopic techniques such as circular dichroism and intrinsic fluorescence is thus very effective in understanding the conformational transitions of proteins and protein complexes.
AB - In order to improve the low signal-to-noise ratio of the time-resolved small-angle X-ray scattering, we have used a two-dimensional X-ray detector with a beryllium-windowed X-ray image intensifier and a charge-coupled device as an image sensor, and applied this to studies on (1) the kinetic folding reaction of α-lactalbumin, which accumulates the molten globule-like intermediate at an early stage of refolding and (2) the cooperative conformational transition of Escherichia coli chaperonin GroEL induced by ATP, which occurs in an allosteric manner between the close and open conformational states. In the α-lactalbumin reaction, we have firmly established the equivalence between the kinetic intermediate and the equilibrium molten globule state, and obtained further information about dehydration from the highly hydrated folding intermediate during a late stage of refolding. In the chaperonin study, we have successfully observed the kinetics of the allosteric transition of GroEL that occurs with a rate constant of about 3-4 s-1 at 5°C. The combination of the time-resolved X-ray scattering with other spectroscopic techniques such as circular dichroism and intrinsic fluorescence is thus very effective in understanding the conformational transitions of proteins and protein complexes.
UR - http://www.scopus.com/inward/record.url?scp=0036965963&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036965963&partnerID=8YFLogxK
U2 - 10.1155/2002/428646
DO - 10.1155/2002/428646
M3 - Article
AN - SCOPUS:0036965963
VL - 16
SP - 127
EP - 138
JO - Spectroscopy (New York)
JF - Spectroscopy (New York)
SN - 0712-4813
IS - 3-4
ER -