Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
11
|
| pubmed:dateCreated |
1990-12-11
|
| pubmed:abstractText |
A fully encoding cDNA for the high-molecular-weight rat neurofilament protein (NF-H) has been isolated from a lambda gt11 library, sequenced and subcloned into eukaryotic expression vectors. Sequence analysis shows that rat NF-H has an overall homology of 72 and 88% with human and mouse NF-H, respectively. The head and rod domains are almost entirely identical, and the divergences are due to differences in the long C-terminal extensions of the molecule. The consensus phosphorylation sequence for neurofilaments Lys-Ser-Pro (KSP) is present 52 times. The predicted molecular mass of the protein is 115 kDa, 42% lower than that observed by SDS-PAGE. Upon transfection into vimentin-containing fibroblasts, such as L tk-, L929, and 3T6 cells, NF-H is seen distributed with vimentin by light and electron microscopic examinations indicating that copolymers of NF-H and vimentin are formed in these cells. Only a negligible proportion of the cells is positive when stained with a number of antibodies directed against phosphorylated NF-H epitopes. This is in contrast with the middle molecular weight NF protein (NF-M) transfected into L tk- and L929 cells, which can readily be detected by antibodies against phosphorylated neurofilament epitopes. The mobilities of the transfected protein on 1- and 2-dimensional gels confirm that NF-H is predominantly in a nonphosphorylated form. These results indicate that phosphorylation of NF-H, but not NF-M, on the KSP sequence is due to protein kinases, which are not present in fibroblasts and are presumably NF-H specific. The stable NF-H-expressing cell lines can therefore be used to study these putative neurofilament kinases in vitro and in vivo.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Intermediate Filament Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Neurofilament Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Vimentin,
http://linkedlifedata.com/resource/pubmed/chemical/neurofilament protein H
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0270-6474
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
10
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
3714-26
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:2230956-Amino Acid Sequence,
pubmed-meshheading:2230956-Animals,
pubmed-meshheading:2230956-Base Sequence,
pubmed-meshheading:2230956-Brain,
pubmed-meshheading:2230956-Cloning, Molecular,
pubmed-meshheading:2230956-DNA,
pubmed-meshheading:2230956-Fluorescent Antibody Technique,
pubmed-meshheading:2230956-Gene Library,
pubmed-meshheading:2230956-Humans,
pubmed-meshheading:2230956-Intermediate Filament Proteins,
pubmed-meshheading:2230956-Intermediate Filaments,
pubmed-meshheading:2230956-L Cells (Cell Line),
pubmed-meshheading:2230956-Molecular Sequence Data,
pubmed-meshheading:2230956-Molecular Weight,
pubmed-meshheading:2230956-Neurofilament Proteins,
pubmed-meshheading:2230956-Phosphorylation,
pubmed-meshheading:2230956-Protein Biosynthesis,
pubmed-meshheading:2230956-Rats,
pubmed-meshheading:2230956-Sequence Homology, Nucleic Acid,
pubmed-meshheading:2230956-Transcription, Genetic,
pubmed-meshheading:2230956-Transfection,
pubmed-meshheading:2230956-Vimentin
|
| pubmed:year |
1990
|
| pubmed:articleTitle |
Transfected rat high-molecular-weight neurofilament (NF-H) coassembles with vimentin in a predominantly nonphosphorylated form.
|
| pubmed:affiliation |
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
|
| pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.
|