Cellular phosphorylation of neurofilament heavy-chain by cyclin-dependent kinase-5 masks the epitope for monoclonal antibody N52

Autor:	S, Guidato, N P, Bajaj, C C, Miller
Rok vydání:	1996
Předmět:	Neurons Blotting Western Antibodies Monoclonal Cyclin-Dependent Kinase 5 Protein Serine-Threonine Kinases Cyclin-Dependent Kinases Cell Line Rats Molecular Weight Epitopes Neurofilament Proteins Animals Humans Electrophoresis Polyacrylamide Gel Phosphorylation
Zdroj:	Neuroscience letters. 217(2-3)
ISSN:	0304-3940
Popis:	N52 is a widely used monoclonal antibody reported to recognise both phosphorylated and non-phosphorylated forms of neurofilament (NF)-H. N52 is therefore classified as a phosphorylation-independent-type antibody. N52 is strongly reactive with NF-H in COS cells transfected with NF-H alone but co-transfection of NF-H with the neurofilament kinase cdk-5 and one of its activators p35, induced phosphorylation of NF-H that abolished this reactivity. Treatment of the cdk-5 phosphorylated NF-H with alkaline phosphatase so as to remove phosphate restored N52 reactivity. A fragment of NF-H containing the consensus cdk-5 sites was reactive with N52 but following co-transfection with cdk-5/p35 a slower migrating fragment species generated by cdk-5 was not labelled by N52. These results demonstrate that N52 is not a truly phosphorylation-independent-type NF-H antibody and suggest that the N52 epitope contains sites targeted for phosphorylation by cdk-5.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::b2f09716258c6f45d3e21cd02c7842a4 https://pubmed.ncbi.nlm.nih.gov/8916096 Zobrazit plný text záznamu Full Text from ScienceDirect