Asymmetry of the GroEL-GroES complex under physiological conditions as revealed by small-angle x-ray scattering.
Despite the well-known functional importance of GroEL-GroES complex formation during the chaperonin cycle, the stoichiometry of the complex has not been clarified. The complex can occur either as an asymmetric 1:1 GroEL-GroES complex or as a symmetric 1:2 GroEL-GroES complex, although it remains uncertain which type is predominant under physiological conditions. To resolve this question, we studied the structure of the GroEL-GroES complex under physiological conditions by small-angle x-ray scattering, which is a powerful technique to directly observe the structure of the protein complex in solution. We evaluated molecular structural parameters, the radius of gyration and the maximum dimension of the complex, from the x-ray scattering patterns under various nucleotide conditions (3 mM ADP, 3 mM ATP gamma S, and 3 mM ATP in 10 mM MgCl(2) and 100 mM KCl) at three different temperatures (10 degrees C, 25 degrees C, and 37 degrees C). We then compared the experimentally observed scattering patterns with those calculated from the known x-ray crystallographic structures of the GroEL-GroES complex. The results clearly demonstrated that the asymmetric complex must be the major species stably present in solution under physiological conditions. On the other hand, in the presence of ATP (3 mM) and beryllium fluoride (10 mM NaF and 300 microM BeCl(2)), we observed the formation of a stable symmetric complex, suggesting the existence of a transiently formed symmetric complex during the chaperonin cycle.
Inobe
Tomonao
T
Department of Physics, School of Science, University of Tokyo and CREST, Japan Science and Technology Agency, Tokyo 113-0033, Japan.
Takahashi
Kazunobu
K
Maki
Kosuke
K
Enoki
Sawako
S
Kamagata
Kiyoto
K
Kadooka
Akio
A
Arai
Munehito
M
Kuwajima
Kunihiro
K
D001665:Binding Sites; D018835:Chaperonin 10 / Q000737:chemistry* / Q000648:ultrastructure*; D018834:Chaperonin 60 / Q000737:chemistry* / Q000648:ultrastructure*; D003198:Computer Simulation; D008956:Models, Chemical; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure; D011485:Protein Binding; D011487:Protein Conformation; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction / Q000379:methods*
D018835:Chaperonin 10; D018834:Chaperonin 60; D046912:Multiprotein Complexes