Structural changes of the Escherichia coli GroEL-GroES chaperonins upon complex formation in solution: a neutron small angle scattering study.
We applied neutron scattering in conjunction with deuterium (D-) labeling in order to obtain information about the domain structure of GroEL and GroES isolated and in the complex. Each subunit of the heptameric GroES consists of two domains, a core domain (Met1 to Lys15 and Lys34 to Ala97) and an intervening loop region (Glu16 to Ala33). Neutron scattering shows that both regions change their conformation upon GroEL/GroES complex formation. The interdomain angle between the core regions of the heptameric GroES increases from 120 to 140 degrees, leading to a less dome-like shape of GroES, and the loop regions turn inwards by 75 degrees. The 23 C-terminal amino acids of the 14 GroEL subunits (Lys526 to Met548), which are unresolved in the crystal structure, are located either at the bottom of the cavity formed by the seven-membered GroEL ring or at the inner wall of the cavity. Upon complex formation the apical domains of GroEL move outwards, which facilitates binding of GroES at a Gro-EL-GroES center-to-center distance of (87 +/- 8) A. These structural changes may be important for the dissociation of the unfolded protein bound to the central cavity upon GroES binding. The overall structure determined by neutron scattering in solution tallies with the crystallographic model published after completion of this study. Differences in the conformation of GroES observed in the complex by the two methods support the view that the chaperonin complex is a flexible molecule which might switch in solution between different conformations.
Stegmann
R
R
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Manakova
E
E
Rössle
M
M
Heumann
H
H
Nieba-Axmann
S E
SE
Plückthun
A
A
Hermann
T
T
May
R P
RP
Wiedenmann
A
A
D018835:Chaperonin 10 / Q000737:chemistry*; D018834:Chaperonin 60 / Q000737:chemistry*; D003903:Deuterium; D004926:Escherichia coli / Q000737:chemistry; D046911:Macromolecular Substances; D008958:Models, Molecular; D009502:Neutrons; D011487:Protein Conformation; D012542:Scattering, Radiation; D012996:Solutions
D018835:Chaperonin 10; D018834:Chaperonin 60; D046911:Macromolecular Substances; D012996:Solutions; D003903:Deuterium