Effects of urea and guanidine hydrochloride on the sliding
movement of actin filaments with ATP hydrolysis by myosin molecules
Ryusei Kumemoto, Yoshitsugu
Hosogoe, Naoki Nomura, and Kuniyuki Hatori*
Department of Bio-System
Engineering,
Graduate School of Science and
Technology,
Yamagata University
Yonezawa 992-8510, Japan
Summary
@ To evaluate the role of the hydration layer
on the protein surface of actomyosin, we compared the effects of urea and
guanidine-HCl on the sliding velocities and ATPase activities of the
actin-heavy meromyosin (HMM) system. Both chemicals denature proteins, but only
urea perturbs the hydration layer. Both the sliding velocity of actin filaments
and actin-activated ATPase activity decreased with increasing urea concentrations.
The sliding movement was completely inhibited at 1.0 M urea, while actin
filaments were bound to HMM molecules fixed on the glass surface. Guanidine-HCl
(0–0.05 M) drastically decreased both the sliding velocity and ATPase
activation of acto-HMM complexes. Under this
condition, actin filaments almost detached from HMM molecules. In contrast, the
ATPase activity of HMM without actin filaments was almost independent of urea
concentrations below 1.0 M and guanidine-HCl concentrations below 0.05 M. An increase
in urea concentrations up to 2.0 M partly induced changes in the ternary
structure of HMM molecules, while the actin filaments were stable in this
concentration range. Hydration changes around such actomyosin complexes may
alter both the stability of part of the myosin molecules, and the affinity for
force transmission between actin filaments and myosin heads.
Keywords:
actomyosin; ATP hydrolysis; denaturant; hydration; motility