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Journal of Cell Science, Vol 106, Issue 4 1283-1290, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

Immunological properties and cDNA sequence analysis of an intermediate-filament-like protein from squid neuronal tissue

J Adjaye, PJ Marsh and PA Eagles
Department of Molecular Biology and Biophysics, Randall Institute, King's College London, UK.

A cDNA library has been constructed in the expression vector lambda gt11 from mRNA isolated from squid (Loligo forbesi) optic lobes. The library was screened with antibodies generated against purified squid neurofilaments. A positive clone was isolated, which harboured a lambda gt11 recombinant having an insert size of 3.5 kb. Hybridization analysis by Southern and northern blotting showed that the corresponding protein is encoded by a single gene that gives rise to a transcript of 2.6 kb. Translation of the full nucleotide sequence of the gene revealed an open reading frame covering 557 amino acids. This squid-neurofilament-like protein, SNLK, bears the characteristic N-terminal head, rod and C-terminal tail domains present in all intermediate filament (IF) proteins. The rod has the classical heptad repeats indicating coiled-coil-forming ability, and the predicted lengths of the coils are similar to coils 1a, 1b and 2 of intermediate filaments. At the C-terminal end of the rod there is a strongly conserved IF epitope, and a fusion protein containing SNLK is recognised by the pan-specific intermediate filament antibody, IFA. A polyclonal antibody raised against SNLK has been used to show that the protein is present only in neuronal tissues and that it is immunologically related to neurofilaments from Myxicola but not from mammals.
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This article has been cited by other articles:


Home page
 J. Cell Sci.Home page
J Adjaye, U Plessmann, K Weber, and H Dodemont
Characterisation of neurofilament protein NF70 mRNA from the gastropod Helix aspersa reveals that neuronal and non-neuronal intermediate filament proteins of cerebral ganglia arise from separate lamin-related genes
J. Cell Sci., January 11, 1995; 108(11): 3581 - 3590.
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© The Company of Biologists Ltd 1993