Abstract

The long term goal of this project is to characterize the structure and function of P-57, a novel calmodulin (CaM) binding protein from brain. We have recently purified P-57 to apparent homogeneity from bovine cerebral cortex membranes. In contrast to other CaM binding proteins, P-57 apparently has higher affinity for CaM in the absence of bound Ca2+ than in its presence and Ca2+ causes the release of CaM from P-57. It is hypothesized that P-57 may function to bind CaM at some local site within the cell and release CaM in response to increases in free Ca2+.
The specific aims of this project include quantitation of binding between P-57 and CaM in the presence and absence of Ca2+, determining if P-57 interacts with other polypeptides in brain, examination of the tissue distribution of P-57, characterization of the physical properties of P-57, characterization of cAMP dependent protein kinase catalyzed phosphorylation of P-57, and determination of the influence of phosphorylation on the affinity of P-57 for CaM. Homogeneous P-57 in mg quantities, anti-P57 rabbit polyclonal antibodies, and fluorescent labeled CaM derivatives are currently available for the proposed experiments.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033708-02
Application #
3283630
Study Section
Biochemistry Study Section (BIO)
Project Start
1984-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Liu, Y; Fisher, D A; Storm, D R (1994) Intracellular sorting of neuromodulin (GAP-43) mutants modified in the membrane targeting domain. J Neurosci 14:5807-17
Liu, Y; Fisher, D A; Storm, D R (1993) Analysis of the palmitoylation and membrane targeting domain of neuromodulin (GAP-43) by site-specific mutagenesis. Biochemistry 32:10714-9
Chapman, E R; Alexander, K; Vorherr, T et al. (1992) Fluorescence energy transfer analysis of calmodulin-peptide complexes. Biochemistry 31:12819-25
Chapman, E R; Estep, R P; Storm, D R (1992) Palmitylation of neuromodulin (GAP-43) is not required for phosphorylation by protein kinase C. J Biol Chem 267:25233-8
Chapman, E R; Au, D; Alexander, K A et al. (1991) Characterization of the calmodulin binding domain of neuromodulin. Functional significance of serine 41 and phenylalanine 42. J Biol Chem 266:207-13
Apel, E D; Litchfield, D W; Clark, R H et al. (1991) Phosphorylation of neuromodulin (GAP-43) by casein kinase II. Identification of phosphorylation sites and regulation by calmodulin. J Biol Chem 266:10544-51
Chapman, E R; Au, D; Nicolson, T A et al. (1991) Mutagenesis of the calmodulin binding domain of neuromodulin. Prog Brain Res 89:37-44
Liu, Y C; Chapman, E R; Storm, D R (1991) Targeting of neuromodulin (GAP-43) fusion proteins to growth cones in cultured rat embryonic neurons. Neuron 6:411-20
Liu, Y C; Storm, D R (1991) Expression of a neuromodulin-beta-galactosidase fusion protein in primary cultured neurons and its accumulation in growth cones. Mol Cell Biochem 104:29-34
Apel, E D; Byford, M F; Au, D et al. (1990) Identification of the protein kinase C phosphorylation site in neuromodulin. Biochemistry 29:2330-5

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