The RII subunit of cAMP-dependent protein kinase binds to a common amino-terminal domain in microtubule-associated proteins 2A, 2B, and 2C.
Three products of the MAP2 gene are known: MAP2A and MAP2B (Mr approximately 200,000) and MAP2C (Mr 70,000). The structural relationship between these MAPs and the basis for their diversity in size are unknown. Previously, we found that a significant fraction of type II cAMP-dependent protein kinase was associated via its regulatory subunits with MAP2A and MAP2B. We now use an antibody prepared against the microtubule binding domain of MAP2A and MAP2B to identify MAP2C. All three forms of MAP2 bound to cAMP affinity columns and reacted with 32P-labeled RII in a blot overlay assay. By assaying proteolytic fragments of MAP2A and MAP2B as well as segments of MAP2 expressed in E. coli, the binding site for RII was localized to an 83 amino acid stretch at the distal (amino-terminal) end of the MAP2 arm domain. Therefore, the microtubule binding and RII binding domains are located at extreme opposite ends of MAP2A and MAP2B, and both are conserved in the much shorter MAP2C.
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Cell Biology and Neurobiology Groups, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Dingus
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Bayley
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Vallee
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D000818:Animals; D000906:Antibodies / Q000276:immunology; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D003429:Cross Reactions; D000242:Cyclic AMP / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
D000906:Antibodies; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins; C015482:protein kinase modulator; D000242:Cyclic AMP; D011494:Protein Kinases