Calcium and calmodulin inhibit phosphorylation of a novel auditory nerve protein.
The growing use of cochlear prosthetic devices and demonstrations of direct ototoxic insult to spiral ganglion neurons make it imperative to gain an understanding of intracellular biochemical regulation in primary sensory neurons. Calcium and calmodulin regulate many aspects of neuronal cellular physiology through stimulation of protein kinase activity. We have previously demonstrated the presence of calmodulin-dependent protein kinase substrates in the guinea pig modiolus and, additionally, the presence of two proteins (12 kDa and 81 kDa, designated as p12 and p81) whose phosphorylation is blocked by calcium and calmodulin (Coling and Schacht, 1991). Here, we investigate three models for this unusual regulatory mechanism. The effects of calcium, calmodulin and trifluoperazine on dephosphorylation of both proteins suggests that calmodulin inhibits protein kinase activity. P81 was identified by immunoprecipitation as the myristoylated alanine-rich C kinase substrate (MARCKS), a ubiquitous actin-binding protein. Two observations indicate that MARCKS may be regulated differently in acoustic nerve than in cerebral cortex. 32P incorporation was significantly higher in acoustic nerve than in brain. The calmodulin-dependent block of MARCKS phosphorylation was observed only in acoustic nerve. p12 shares several characteristics with myelin basic protein (MBP). We used a double label assay with 32P autoradiography and immunoblotting to show that p12 is in fact distinct from MBP. We suggest that either p12 or p12 kinase may be either specific to the peripheral auditory system or novel marker proteins for that tissue.
Coling
D E
DE
Kresge Hearing Research Institute, University of Michigan, Ann Arbor 48109-0506.
Naik
R M
RM
Schacht
J
J
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002147:Calmodulin / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000737:chemistry / Q000378:metabolism; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D011506:Proteins / Q000378:metabolism*; D013136:Spiral Ganglion / Q000187:drug effects / Q000378:metabolism; D014268:Trifluoperazine / Q000494:pharmacology*; D000159:Vestibulocochlear Nerve / Q000378:metabolism*
D002147:Calmodulin; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; D047428:Protein Kinase Inhibitors; D011506:Proteins; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D014268:Trifluoperazine; D011493:Protein Kinase C; D002118:Calcium