Neuron
Volume 3, Issue 5, November 1989, Pages 631-638
Journal home page for Neuron

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

https://doi.org/10.1016/0896-6273(89)90273-0Get rights and content

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.

References (53)

  • E. Nishida et al.

    Tyrosine phosphorylation by the epidermal growth factor receptor kinase induces functional alterations in microtubule-associated protein 2

    J. Biol. Chem.

    (1987)
  • C.S. Rubin et al.

    Characterization and comparison of membrane-associated and cytosolic CAMP-dependent protein kinases. Physicochemical and immunological studies on bovine cerebral cortex protein kinases

    J. Biol. Chem.

    (1979)
  • C.S. Rubin et al.

    Posttranslational Covalent Modification of Proteins

  • D. Sarkar et al.

    Identification of a calmodulin binding protein that co-purifies with the regulatory subunit of brain protein kinase 11

    J. Biol. Chem.

    (1984)
  • J.C. Stein et al.

    Differential expression of isoforms of the regulatory subunit of type II cAMP-dependent protein kinase in rat neurons, astrocytes, and oligodendrocytes

    J. Biol. Chem.

    (1987)
  • W.E. Theurkauf et al.

    Molecular characterization of the cAMP-dependent protein kinase bound to microtubuleassociated protein 2

    J. Biol. Chem.

    (1982)
  • W.E. Theurkauf et al.

    Extensive cAMP-dependent and cAMP-independent phosphorylation of microtubule-associated protein 2

    J. Biol. Chem.

    (1983)
  • S. Tsuyama et al.

    Numerous phosphates of microtubule-associated protein 2 in living rat brain

    J. Biol. Chem.

    (1987)
  • J. Vieira et al.

    The pUC plasmids, an M13mp7derived system for insertion mutagenesis and sequencing with synthetic universal primers

    Gene

    (1982)
  • S.L. Weldon et al.

    The regulatory subunit of neural cAMP-dependent protein kinase II represents a unique gene product

    J. Biol. Chem.

    (1985)
  • P.J. Blackshear et al.

    Protein kinases 1988: a current perspective

    FASEB J.

    (1988)
  • P.Y. Chou et al.

    Prediction of protein conformation

    Biochemistry

    (1974)
  • M. Dammerman et al.

    Isolation and characterization of cDNA clones encoding epitopes shared with Alzheimer neurofibrillarytangles

    J. Neurosci. Res.

    (1988)
  • M. Dammerman et al.

    Sequence of a human MAP-2 region sharing epitopes with Alzheimer neurofibrillary tangles

    J. Neurosci. Res.

    (1989)
  • R. De Camilli et al.

    Distribution of microtubule-associated protein 2 in the nervous system of the rat studied by immunofluorescence

    Neuroscience

    (1984)
  • P. De Camilli et al.

    Heterogeneous distribution of the cAMP receptor protein RII in the nervous system: evidence for its intracellular accumulation on microtubules, microtubule-organizing centers, and in the area of the Golgi complex

    J. Cell Biol.

    (1986)
  • Cited by (120)

    • Specific phosphorylation of microtubule-associated protein 2c by extracellular signal–regulated kinase reduces interactions at its Pro-rich regions

      2022, Journal of Biological Chemistry
      Citation Excerpt :

      Despite its disordered nature, MAP2c was identified as one of the AKAPs. Residues 80 to 120 in the N-terminal domain of MAP2c bind RIIα-PKA (54) and phosphorylation of Ser136 has been suggested to interfere with its binding (36). Our direct monitoring of RIIα-PKA binding to MAP2c revealed that RIIα-PKA is indeed recognized by residues 80 to 120 of MAP2c, but motions of many residues throughout the whole MAP2c sequence are restricted when the complex is formed (blue circles in Fig. 4A).

    • Changes in microtubule turnover accompany synaptic plasticity and memory formation in response to contextual fear conditioning in mice

      2010, Neuroscience
      Citation Excerpt :

      These include a high level of assembly and dependence of MAP2-associated MTs on signaling via the cAMP/PKA pathway. Interestingly, the regulatory subunit of the PKA has been reported to bind to the amino termini of the MAP2 isoforms (Rubino et al., 1989). Thus newly assembled MAP2-associated MTs might stimulate further recruitment of PKA as part of memory formation; conversely, cAMP might promote the growth of MAP2-associated MTs through activation of locally recruited PKA.

    • Selectivity in enrichment of cAMP-dependent protein kinase regulatory subunits type I and type II and their interactors using modified cAMP affinity resins

      2009, Molecular and Cellular Proteomics
      Citation Excerpt :

      The PKA isoforms, RIIα and RIIβ (white bars on the left) show a log 2 ratio of 3.2 (stdev = 0.12) and 3.9 (stdev = 0.17) whereas RIα and RIβ have similar log 2 ratios of −0.79 (stdev = 0.20) and −0.89 (stdev = 0.16), respectively. Furthermore, 6 AKAPs have similar ratios compared with the RII isoforms:AKAP5, AKAP7, AKAP9, AKAP11, MAP2, and AKAP14 (dark gray bars), indicating they are RII specific AKAPs, in agreement with literature data for AKAP5 (25), AKAP7 (26), AKAP11 (27), and MAP2 (28). On the other hand, AKAP1 (light gray bars) had average ratios of around one indicating that it has similar binding affinities to both RI and RII, so it can be classified as dual specificity AKAP.

    View all citing articles on Scopus

    Present Address: Department of Biochemical Genetics and Metabolism, Rockefeller University, New York, New York 10021.

    View full text