Binding of Fyn to MAP-2c through an SH3 binding domain. Regulation of the interaction by ERK2

S P Zamora-Leon; G Lee; P Davies; B Shafit-Zagardo

doi:10.1074/jbc.M107807200

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Binding of Fyn to MAP-2c through an SH3 binding domain. Regulation of the interaction by ERK2

Journal article

Open access

Peer reviewed

Binding of Fyn to MAP-2c through an SH3 binding domain. Regulation of the interaction by ERK2

S P Zamora-Leon, G Lee, P Davies and B Shafit-Zagardo

The Journal of biological chemistry, Vol.276(43), pp.39950-39958

10/26/2001

DOI: 10.1074/jbc.M107807200

PMID: 11546790

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Abstract

Microtubule-associated protein 2 (MAP-2) isoforms are developmentally expressed in the nervous system and contain a number of functional domains. Adjacent to the first repeat of the microtubule-binding domain is an RTPPKSP motif for binding SH3 domains. To identify SH3-containing proteins that interact with MAP-2, transfections, filter overlay assays, glutathione S-transferase (GST)-mediated binding assays, co-immunoprecipitations and enzyme-linked immunosorbent assays were performed. Transfections of MAP-2a, MAP-2b, and MAP-2c constructs into COS7 cells, followed by incubation of the cell lysates with SH3-GST fusion proteins, determined that the strongest interaction was between MAP-2c and the non-receptor tyrosine kinase Fyn; however, MAP-2b and MAP-2c also bound to Grb2. Co-immunoprecipitation of Fyn and MAP-2c from human fetal homogenates confirmed the interaction in vivo. MAP-2 synthetic peptides spanning the RTPPKSP motif bound to Fyn, and the interaction was regulated by phosphorylation. Co-transfections with MAP-2c and the extracellular signal-regulated kinase 2 (ERK2) demonstrated that MAP-2c is threonine/serine-phosphorylated on its RTPPKSP motif and that threonine phosphorylation abolished the MAP-2c/Fyn binding. Kinase assays and co-transfection of MAP-2c and Fyn confirmed that Fyn tyrosine kinase phosphorylates MAP-2c. Thus, the activation of signaling pathways may regulate cytoskeletal dynamics by altering the state of phosphorylation of MAP-2 by both ERK2 and Fyn kinase.

Brain - embryology

Phosphorylation

Microtubule-Associated Proteins - genetics

Microtubule-Associated Proteins - metabolism

Protein-Tyrosine Kinases - metabolism

Humans

Phosphoprotein Phosphatases - metabolism

Proto-Oncogene Proteins c-fyn

Molecular Sequence Data

Phosphoproteins - metabolism

Threonine - metabolism

Brain - metabolism

Protein-Tyrosine Kinases - genetics

Protein Isoforms - metabolism

Binding Sites

Protein Structure, Tertiary

Proto-Oncogene Proteins - metabolism

Recombinant Proteins - metabolism

Amino Acid Sequence

Glutathione Transferase - metabolism

Proto-Oncogene Proteins - genetics

src Homology Domains

Adaptor Proteins, Signal Transducing

Phosphoprotein Phosphatases - genetics

Proteins - metabolism

GRB2 Adaptor Protein

Models, Biological

Protein Binding

Cytoskeleton - physiology

Mitogen-Activated Protein Kinase 1 - metabolism

Details

Title: Subtitle: Binding of Fyn to MAP-2c through an SH3 binding domain. Regulation of the interaction by ERK2
Creators: S P Zamora-Leon - Department of Pathology, Albert Einstein College of Medicine, Bronx, NY 10461, USA
G Lee
P Davies
B Shafit-Zagardo
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.276(43), pp.39950-39958
DOI: 10.1074/jbc.M107807200
PMID: 11546790
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: MH 38623 / NIMH NIH HHS NS 38102 / NINDS NIH HHS NS 32100 / NINDS NIH HHS
Language: English
Date published: 10/26/2001
Academic Unit: Iowa Neuroscience Institute; Immunology; Internal Medicine
Record Identifier: 9984065400202771

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