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An Immunoblot Study of Neurofilament Degradation In Situ and During Calcium‐Activated Proteolysis
Author(s) -
Schlaepfer W. W.,
Lee C.,
Lee V. M.Y.,
Zimmerman U.J. P.
Publication year - 1985
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1985.tb05442.x
Subject(s) - proteolysis , neurofilament , calcium , calpain , polyclonal antibodies , microbiology and biotechnology , protein degradation , chemistry , calcium binding protein , fragmentation (computing) , in vivo , biochemistry , biology , immunohistochemistry , antibody , immunology , enzyme , ecology , organic chemistry
The degradation of neurofilament (NF) proteins was examined by immunoblot methods to identify, characterize, and monitor the appearance of immunoreactive breakdown products during the loss of NF triplet proteins. Individual NF proteins and their breakdown products were identified using polyclonal and monoclonal antibodies to NF proteins. NF degradation was compared during calcium‐activated proteolysis of isolated rat NF, during an experimental influx of calcium into excised rat spinal nerve roots, and during NF breakdown in transected rat peripheral nerve. These different experimental conditions produced similar patterns of NF fragmentation, including the transient appearance of NF immunobands between M r 150,000–200,000 and 110,000–120,000 as well as the appearance and accumulation of NF immunobands between M r 45,000 and 65,000. Most immunoreactive NF fragments remained Triton‐insoluble. Low levels of the same immunoreactive fragments were present in control neural tissues, suggesting that calciumactivated proteolysis may be operative in the turnover and/or processing of NF proteins in vivo. Very similar patterns of NF degradation during experimental calcium influxes into different CNS and PNS tissues are indicative of the widespread distribution of calcium‐activated NF protease in neural tissues.