PMID- 9148897
OWN - NLM
STAT- MEDLINE
DCOM- 19970619
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 272
IP  - 20
DP  - 1997 May 16
TI  - Identification of phosphorylation sites on neurofilament proteins by 
      nanoelectrospray mass spectrometry.
PG  - 12922-7
AB  - Neurofilament (NF) proteins are intermediate filaments found in the neuronal 
      cytoskeleton. Phosphorylation of these proteins is considered an important factor 
      in the assembly of filaments and determination of filament caliber and stability. 
      Mammalian neurofilaments are composed of three polypeptide subunits, NF-L, NF-M, 
      and NF-H, all of which are phosphorylated. Here we used techniques for the mass 
      spectrometric sequencing of proteins from polyacrylamide gels to analyze in vivo 
      phosphorylation sites on NF-M and NF-L. Neurofilaments were isolated from rat 
      brain and enzymatically digested in gel. The resulting peptides were analyzed and 
      sequence data obtained by nanoelectrospray mass spectrometry. Four 
      phosphorylation sites have been found in the C-terminal domain of NF-M: serines 
      603, 608, 666, and 766. Two of these are found in lysine-serine-proline (KSP) 
      motifs and two in the variant motifs, glutamic acid-serine-proline (ESP) and 
      valine-serine-proline (VSP). Serine 55 in NF-L was not found to be 
      phosphorylated, which confirms the possible role of phosphorylation and 
      dephosphorylation of this site in early neurofilament assembly. The techniques 
      used enable sequence data and characterization of posttranslational modifications 
      to be obtained for each individual subunit directly from polyacrylamide gels.
FAU - Betts, J C
AU  - Betts JC
AD  - Department of Neuroscience, The Institute of Psychiatry, De Crespigny Park, 
      London, SE5 8AF, United Kingdom. wxx35951@ggr.co.uk
FAU - Blackstock, W P
AU  - Blackstock WP
FAU - Ward, M A
AU  - Ward MA
FAU - Anderton, B H
AU  - Anderton BH
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Neurofilament Proteins)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - *Brain Chemistry
MH  - Mass Spectrometry
MH  - Molecular Sequence Data
MH  - Neurofilament Proteins/*chemistry
MH  - Phosphorylation
MH  - Rats
EDAT- 1997/05/16 00:00
MHDA- 1997/05/16 00:01
CRDT- 1997/05/16 00:00
PHST- 1997/05/16 00:00 [pubmed]
PHST- 1997/05/16 00:01 [medline]
PHST- 1997/05/16 00:00 [entrez]
AID - S0021-9258(19)56875-2 [pii]
AID - 10.1074/jbc.272.20.12922 [doi]
PST - ppublish
SO  - J Biol Chem. 1997 May 16;272(20):12922-7. doi: 10.1074/jbc.272.20.12922.