Ca2+-mediated phosphorylation and proteolysis activity associated with the cytoskeletal fraction from cerebral cortex of rats

被引:4
作者
deFreitas, MS [1 ]
deMattosDutra, A [1 ]
Wannmacher, CMD [1 ]
PessoaPureur, R [1 ]
机构
[1] UNIV FED RIO GRANDE SUL,INST BIOCIENCIAS,DEPT BIOQUIM,BR-90046900 PORTO ALEGRE,RS,BRAZIL
关键词
cytoskeletal proteins; phosphorylation; proteolysis; cerebral cortex;
D O I
10.1007/BF02533096
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We describe a Triton-insoluble cytoskeletal fraction extracted from cerebral cortex of young rats retaining an endogenous Ca2+-mediated mechanism acting in vitro on Ca2+/calmodulin-dependent protein kinase II (CaM-KII) activity and on phosphorylation and proteolysis of the 150 kDa neurofilament subunit (NF-M), alpha and beta tubulin. Exogenous Ca2+ induced a 70% decrease in the in vitro phosphorylation of the NF-M and tubulins and a 30-50% decrease in the total amount of these proteins. However, when calpastatin was added basal phosphorylation and NF-M and tubulin content were recovered. Furthermore, exogenous Ca2+/calmodulin induced increased in vitro phosphorylation of the cytoskeletal proteins and CaM-KII activity only in the presence of calpastatin, suggesting the presence of Ca2+-induced calpain-mediated proteolysis. This fraction could be an interesting model to further studies concerning the in vitro effects of Ca2+-mediated protein kinases and proteases associated with the cytoskeletal fraction.
引用
收藏
页码:1489 / 1495
页数:7
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