Innervation-dependent phosphorylation and accumulation of alphaB-crystallin and Hsp27 as insoluble complexes in disused muscle.
Levels and phosphorylation states of the two small molecular chaperones, alphaB-crystallin and Hsp27, in disused rat soleus muscles were determined by Western blot analysis of extracts with antibodies recognizing each of the two proteins and their phosphorylated serine residues. Increased phosphorylation and relocalization to insoluble fractions were found within a few days of hind-limb suspension. High phosphorylation of alphaB-crystallin at Ser-59 (and to a certain extent, at Ser-45) and of Hsp27 at Ser-15 and Ser-85, along with phosphorylated, active states of p38 and p44/42 mitogen-activated protein kinases were maintained during hind-limb suspension but promptly returned to control levels within a 5-day recovery period. These results are similar to those observed with U373 MG glioma cells exposed to proteasome inhibitors (16). However, the responses of alphaB-crystallin and Hsp27 in suspended soleus muscles did not appear with ipsilateral transection of the sciatic nerve trunk, indicating mediation by nerve activity. The fact that ubiquitinated proteins accumulated in the insoluble fractions of suspended soleus muscle further suggests participation of alphaB-crystallin and Hsp27 in quality control of proteins in disused soleus muscle, with involvement of nerve activity-dependent processes.
Kato
Kanefusa
K
Department of Biochemistry, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi 480-0392, Japan. kato@inst-hsc.pref.aichi.jp
Ito
Hidenori
H
Kamei
Keiko
K
Iwamoto
Ikuko
I
Inaguma
Yutaka
Y
D000818:Animals; D003459:Crystallins / Q000378:metabolism*; D003714:Denervation; D006360:Heat-Shock Proteins / Q000378:metabolism*; D019417:Hindlimb Suspension; D020935:MAP Kinase Signaling System; D046911:Macromolecular Substances; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000294:innervation* / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D012995:Solubility
D003459:Crystallins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances