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
N1 - Funding Information:
We are grateful to Ms. Chris Hubertus for preparation of this manuscript and Mr. Fred Brown for photographic expertise. We acknowledge Dr. Shu-Hui Yen for helpful discussion during the preparation of this manuscript. This work was supported by grants AC-06803 and DK-27736 from the National Institutes Health and NP-377E from the American Cancer Society.
PY - 1989/11
Y1 - 1989/11
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.
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U2 - 10.1016/0896-6273(89)90273-0
DO - 10.1016/0896-6273(89)90273-0
M3 - Article
C2 - 2701845
AN - SCOPUS:0024763898
SN - 0896-6273
VL - 3
SP - 631
EP - 638
JO - Neuron
JF - Neuron
IS - 5
ER -