Characterization of the bovine neurofilament NF-M protein and cDNA sequence, and identification of in vitro and in vivo calpain cleavage sites

Gerry Shaw, Cui Yang, Lu Zhang, Petra Cook, Brian Pike, William D Hill

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Neurofilaments are the major components of the neuronal cytoskeleton, and accumulations of these proteins are associated with several important human diseases. Here we report the cloning and sequencing of bovine NF-M, the first NF-M cloned from a large domestic mammal. The bovine sequence proves to be generally more similar to that of human NF-M than the previously described mammalian sequences, suggesting that bovine neurofilaments are a useful model for biochemical studies of application to humans. However, we noted some unusual features within the 16 lys-ser-pro (KSP2) type phosphopeptide repeats and also note that the number of these repeats correlates well with the size of animal. We also characterized two in vitro calpain cleavage sites by direct peptide sequencing, finding that both are located in the glutamic acid rich E segment. Finally, we show biochemically that the more abundant and stable of these calpain fragments can also be detected in vivo.

Original languageEnglish (US)
Pages (from-to)619-625
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume325
Issue number2
DOIs
StatePublished - Dec 10 2004

Fingerprint

Calpain
Intermediate Filaments
Complementary DNA
Phosphopeptides
Mammals
Cloning
Glutamic Acid
Animals
Cytoskeleton
Peptides
Organism Cloning
Proteins
neurofilament protein M
In Vitro Techniques

Keywords

  • Calpain
  • Cytoskeleton
  • NF-M
  • Neurofilaments
  • Proteolysis
  • cDNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Characterization of the bovine neurofilament NF-M protein and cDNA sequence, and identification of in vitro and in vivo calpain cleavage sites. / Shaw, Gerry; Yang, Cui; Zhang, Lu; Cook, Petra; Pike, Brian; Hill, William D.

In: Biochemical and Biophysical Research Communications, Vol. 325, No. 2, 10.12.2004, p. 619-625.

Research output: Contribution to journalArticle

Shaw, Gerry ; Yang, Cui ; Zhang, Lu ; Cook, Petra ; Pike, Brian ; Hill, William D. / Characterization of the bovine neurofilament NF-M protein and cDNA sequence, and identification of in vitro and in vivo calpain cleavage sites. In: Biochemical and Biophysical Research Communications. 2004 ; Vol. 325, No. 2. pp. 619-625.
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