Effect of alpha-crystallin on thermal aggregation of glycogen phosphorylase b from rabbit skeletal muscle.
Thermal aggregation of rabbit skeletal muscle glycogen phosphorylase b (Phb) has been investigated using dynamic light scattering under conditions of a constant rate of temperature increase (1 K/min). The linear behavior of the dependence of the hydrodynamic radius on temperature for Phb aggregation is consistent with the idea that thermal aggregation of proteins proceeds in the kinetic regime wherein the rate of aggregation is limited by diffusion of the interacting particles (the regime of "diffusion-limited cluster-cluster aggregation"). In the presence of alpha-crystallin, a protein exhibiting chaperone-like activity, the dependence of the hydrodynamic radius on temperature follows the exponential law; this suggests that the aggregation process proceeds in the kinetic regime where the sticking probability for colliding particles becomes lower than unity (the regime of "reaction-limited cluster-cluster aggregation"). Based on analysis of the ratio between the light scattering intensity and the hydrodynamic radius of Phb aggregates, it has been concluded that the addition of alpha-crystallin results in formation of smaller size starting aggregates. The data on differential scanning calorimetry indicate that alpha-crystallin interacts with the intermediates of the unfolding process of the Phb molecule. The proposed scheme of thermal denaturation and aggregation of Phb includes the stage of reversible dissociation of dimers of Phb into monomers, the stage of the formation of the starting aggregates from the denatured monomers of Phb, and the stage of the sticking of the starting aggregates and higher order aggregates. Dissociation of Phb dimer into monomers at elevated temperatures has been confirmed by analytical ultracentrifugation.
Meremyanin
A V
AV
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow 119071, Russia. mer-av@inbi.ras.ru
Eronina
T B
TB
Chebotareva
N A
NA
Kleimenov
S Yu
SY
Yudin
I K
IK
Muranov
K O
KO
Ostrovsky
M A
MA
Kurganov
B I
BI
D000465:Algorithms; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D024981:Glycogen Phosphorylase / Q000737:chemistry*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D018482:Muscle, Skeletal / Q000201:enzymology*; D010763:Phosphorylase b / Q000737:chemistry*; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D011817:Rabbits; D038201:alpha-Crystallins / Q000737:chemistry / Q000494:pharmacology*
D038201:alpha-Crystallins; D024981:Glycogen Phosphorylase; D010763:Phosphorylase b