Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2

Heidi M. Rubino, Marilyn Dammerman, Bridget Shafit-Zagardo, Jack Erlichman

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Microtubule-associated protein 2 (MAP2) binds, and is a substrate for, type 11 cAMP-dependent protein kinase. The structural domain in MAP2 that binds the regulatory subunit (RII) of protein kinase If was identified by expressing fragments of a human MAP2 cDNA in E. coli using the pATH11 vector. Fusion proteins were resolved by SDS-PAGE and transferred to nitrocellulose. The filters were probed with purified bovine heart or brain RII, anti-RII monoclonal antibodies, and 125I-labeled protein A. Binding of RII was localized to a 31 amino acid sequence near the N-terminus of the MAP2 molecule. Fusion proteins containing this fragment bound both heart and brain Rlls in a concentrationdependent manner, but bound heart RII with a higher apparent affinity than brain RII. The amino acid sequence of this fragment (DRETAEEVSARIVQVVTAEAV AVLKGEQEKE) is totally conserved between human and mouse MAP2, suggesting an important role for the RII binding site of MAP2 in neuronal function.

Original languageEnglish (US)
Pages (from-to)631-638
Number of pages8
JournalNeuron
Volume3
Issue number5
DOIs
StatePublished - 1989

Fingerprint

Microtubule-Associated Proteins
Cyclic AMP-Dependent Protein Kinases
Binding Sites
Amino Acid Sequence
Brain
Collodion
Staphylococcal Protein A
Protein Binding
Protein Kinases
Polyacrylamide Gel Electrophoresis
Proteins
Complementary DNA
Monoclonal Antibodies
Escherichia coli
human MAP2 protein

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2. / Rubino, Heidi M.; Dammerman, Marilyn; Shafit-Zagardo, Bridget; Erlichman, Jack.

In: Neuron, Vol. 3, No. 5, 1989, p. 631-638.

Research output: Contribution to journalArticle

@article{eadf4e49d08240928ab999bdd920ad8e,
title = "Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2",
abstract = "Microtubule-associated protein 2 (MAP2) binds, and is a substrate for, type 11 cAMP-dependent protein kinase. The structural domain in MAP2 that binds the regulatory subunit (RII) of protein kinase If was identified by expressing fragments of a human MAP2 cDNA in E. coli using the pATH11 vector. Fusion proteins were resolved by SDS-PAGE and transferred to nitrocellulose. The filters were probed with purified bovine heart or brain RII, anti-RII monoclonal antibodies, and 125I-labeled protein A. Binding of RII was localized to a 31 amino acid sequence near the N-terminus of the MAP2 molecule. Fusion proteins containing this fragment bound both heart and brain Rlls in a concentrationdependent manner, but bound heart RII with a higher apparent affinity than brain RII. The amino acid sequence of this fragment (DRETAEEVSARIVQVVTAEAV AVLKGEQEKE) is totally conserved between human and mouse MAP2, suggesting an important role for the RII binding site of MAP2 in neuronal function.",
author = "Rubino, {Heidi M.} and Marilyn Dammerman and Bridget Shafit-Zagardo and Jack Erlichman",
year = "1989",
doi = "10.1016/0896-6273(89)90273-0",
language = "English (US)",
volume = "3",
pages = "631--638",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "5",

}

TY - JOUR

T1 - Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2

AU - Rubino, Heidi M.

AU - Dammerman, Marilyn

AU - Shafit-Zagardo, Bridget

AU - Erlichman, Jack

PY - 1989

Y1 - 1989

N2 - Microtubule-associated protein 2 (MAP2) binds, and is a substrate for, type 11 cAMP-dependent protein kinase. The structural domain in MAP2 that binds the regulatory subunit (RII) of protein kinase If was identified by expressing fragments of a human MAP2 cDNA in E. coli using the pATH11 vector. Fusion proteins were resolved by SDS-PAGE and transferred to nitrocellulose. The filters were probed with purified bovine heart or brain RII, anti-RII monoclonal antibodies, and 125I-labeled protein A. Binding of RII was localized to a 31 amino acid sequence near the N-terminus of the MAP2 molecule. Fusion proteins containing this fragment bound both heart and brain Rlls in a concentrationdependent manner, but bound heart RII with a higher apparent affinity than brain RII. The amino acid sequence of this fragment (DRETAEEVSARIVQVVTAEAV AVLKGEQEKE) is totally conserved between human and mouse MAP2, suggesting an important role for the RII binding site of MAP2 in neuronal function.

AB - Microtubule-associated protein 2 (MAP2) binds, and is a substrate for, type 11 cAMP-dependent protein kinase. The structural domain in MAP2 that binds the regulatory subunit (RII) of protein kinase If was identified by expressing fragments of a human MAP2 cDNA in E. coli using the pATH11 vector. Fusion proteins were resolved by SDS-PAGE and transferred to nitrocellulose. The filters were probed with purified bovine heart or brain RII, anti-RII monoclonal antibodies, and 125I-labeled protein A. Binding of RII was localized to a 31 amino acid sequence near the N-terminus of the MAP2 molecule. Fusion proteins containing this fragment bound both heart and brain Rlls in a concentrationdependent manner, but bound heart RII with a higher apparent affinity than brain RII. The amino acid sequence of this fragment (DRETAEEVSARIVQVVTAEAV AVLKGEQEKE) is totally conserved between human and mouse MAP2, suggesting an important role for the RII binding site of MAP2 in neuronal function.

UR - http://www.scopus.com/inward/record.url?scp=0024763898&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024763898&partnerID=8YFLogxK

U2 - 10.1016/0896-6273(89)90273-0

DO - 10.1016/0896-6273(89)90273-0

M3 - Article

C2 - 2701845

AN - SCOPUS:0024763898

VL - 3

SP - 631

EP - 638

JO - Neuron

JF - Neuron

SN - 0896-6273

IS - 5

ER -