PMID- 9478962
OWN - NLM
STAT- MEDLINE
DCOM- 19980325
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 273
IP  - 9
DP  - 1998 Feb 27
TI  - Neurofilament (NF) assembly; divergent characteristics of human and rodent NF-L 
      subunits.
PG  - 5101-8
AB  - Previous studies have shown that rodent neurofilaments (NF) are obligate 
      heteropolymers requiring NF-L plus either NF-M or NF-H for filament formation. We 
      have assessed the competence of human NF-L and NF-M to assemble and find that 
      unlike rat NF-L, human NF-L is capable of self-assembly. However, human NF-M 
      cannot form homopolymers and requires the presence of NF-L for incorporation into 
      filaments. To investigate the stage at which filament formation is blocked, the 
      rod domains or the full-length subunits of human NF-L, human NF-M, and rodent 
      NF-L were analyzed in the yeast "interaction trap" system. These studies 
      demonstrated that the fundamental block to filament formation in those 
      neurofilaments that do not form homopolymers is at the level of dimer formation. 
      Based on theoretical biophysical considerations of the requirements for the 
      formation of coiled-coil structures, we predicted which amino acid differences 
      were likely to be responsible for the differing dimerization potentials of the 
      rat and human NF-L rod domains. We tested these predictions using site-specific 
      mutagenesis. Interestingly, single amino acid changes in the rod domains designed 
      to restore or eliminate the coiled-coil propensity were found respectively to 
      convert rat NF-L into a subunit capable of homopolymerization and human NF-L into 
      a protein that is no longer able to self-assemble. Our results additionally 
      suggest that the functional properties of the L12 linker region of human NF-L, 
      generally thought to assume an extended beta-sheet conformation, are consonant 
      with an alpha-helix that positions the heptad repeats before and after it in an 
      orientation that allows coiled-coil dimerization. These studies reveal an 
      important difference between the assembly properties of the human and rodent NF-L 
      subunits possibly suggesting that the initiating events in neurofilament assembly 
      may differ in the two species.
FAU - Carter, J
AU  - Carter J
AD  - Brookdale Center for Developmental and Molecular Biology, Mount Sinai School of 
      Medicine, New York, New York 10029, USA.
FAU - Gragerov, A
AU  - Gragerov A
FAU - Konvicka, K
AU  - Konvicka K
FAU - Elder, G
AU  - Elder G
FAU - Weinstein, H
AU  - Weinstein H
FAU - Lazzarini, R A
AU  - Lazzarini RA
LA  - eng
GR  - DA 00060/DA/NIDA NIH HHS/United States
GR  - DK4 6943/DK/NIDDK NIH HHS/United States
GR  - P50 AGO 5138-11/AG/NIA NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Neurofilament Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (neurofilament protein L)
RN  - 111365-29-8 (neurofilament protein M)
RN  - 30KYC7MIAI (Aspartic Acid)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Aspartic Acid/chemistry
MH  - Dimerization
MH  - Humans
MH  - Intermediate Filaments/*metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Neurofilament Proteins/chemistry/genetics/*metabolism
MH  - Peptide Fragments/chemistry/genetics/metabolism
MH  - Protein Binding
MH  - Protein Structure, Secondary
MH  - Rats
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Repetitive Sequences, Nucleic Acid
MH  - Sequence Homology, Amino Acid
MH  - Species Specificity
MH  - Static Electricity
EDAT- 1998/03/28 00:00
MHDA- 1998/03/28 00:01
CRDT- 1998/03/28 00:00
PHST- 1998/03/28 00:00 [pubmed]
PHST- 1998/03/28 00:01 [medline]
PHST- 1998/03/28 00:00 [entrez]
AID - S0021-9258(17)47086-4 [pii]
AID - 10.1074/jbc.273.9.5101 [doi]
PST - ppublish
SO  - J Biol Chem. 1998 Feb 27;273(9):5101-8. doi: 10.1074/jbc.273.9.5101.