A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin

Peter M Steinert; Ying-Hao Chou; Veena Prahlad; David A D Parry; Lyuben N Marekov; Kenneth C Wu; Shyh-Ing Jang; Robert D Goldman

doi:10.1074/jbc.274.14.9881

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A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin

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A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin

Peter M Steinert, Ying-Hao Chou, Veena Prahlad, David A D Parry, Lyuben N Marekov, Kenneth C Wu, Shyh-Ing Jang and Robert D Goldman

The Journal of biological chemistry, Vol.274(14), pp.9881-9890

04/02/1999

DOI: 10.1074/jbc.274.14.9881

PMID: 10092680

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Abstract

BHK-21 fibroblasts contain type III vimentin/desmin intermediate filament (IF) proteins that typically co-isolate and co-cycle in in vitro experiments with certain high molecular weight proteins. Here, we report purification of one of these and demonstrate that it is in fact the type VI IF protein nestin. Nestin is expressed in several fibroblastic but not epithelioid cell lines. We show that nestin forms homodimers and homotetramers but does not form IF by itself in vitro. In mixtures, nestin preferentially co-assembles with purified vimentin or the type IV IF protein alpha-internexin to form heterodimer coiled-coil molecules. These molecules may co-assemble into 10 nm IF provided that the total amount of nestin does not exceed about 25%. However, nestin does not dimerize with types I/II keratin IF chains. The bulk of the nestin protein consists of a long carboxyl-terminal tail composed of various highly charged peptide repeats. By analogy with the larger neurofilament chains, we postulate that these sequences serve as cross-bridgers or spacers between IF and/or other cytoskeletal constituents. In this way, we propose that direct incorporation of modest amounts of nestin into the backbone of cytoplasmic types III and IV IFs affords a simple yet flexible method for the regulation of their dynamic supramolecular organization and function in cells.

Software

Cell Line

Cricetinae

Nestin

Molecular Weight

Vimentin - metabolism

Molecular Sequence Data

Rats

Nerve Tissue Proteins

Antibodies, Monoclonal

Microscopy, Electron

Sequence Homology, Amino Acid

Kidney - metabolism

Animals

Carrier Proteins - metabolism

Desmin - metabolism

Intermediate Filament Proteins - metabolism

Fibroblasts - metabolism

Intermediate Filament Proteins - chemistry

Details

Title: Subtitle: A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin
Creators: Peter M Steinert - Laboratory of Skin Biology, NIAMS, National Institutes of Health, Bethesda, Maryland 20892-2752, USA. pemast@helix.nih.gov
Ying-Hao Chou
Veena Prahlad
David A D Parry
Lyuben N Marekov
Kenneth C Wu
Shyh-Ing Jang
Robert D Goldman
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.274(14), pp.9881-9890
DOI: 10.1074/jbc.274.14.9881
PMID: 10092680
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM36806-4 / NIGMS NIH HHS
Language: English
Date published: 04/02/1999
Academic Unit: Iowa Neuroscience Institute; Biology
Record Identifier: 9984070717802771

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