Altered phosphorylation of growth-associated protein B50/GAP-43 in Alzheimer disease with high neurofibrillary tangle density.

The growth-associated phosphoprotein B50/GAP-43, associated with axonal proliferation and regeneration, was isolated from superior temporal gyrus (area 22) of seven control and eight Alzheimer disease (AD) postmortem human brains. Membrane and cytoplasmic proteins were fractionated and B50/GAP-43 was isolated by reverse-phase HPLC and gel electrophoresis. B50/GAP-43 was identified with rabbit polyclonal antibodies 4P3 (generated against the calmodulin binding domain of B50/GAP-43) and 1B5 (generated against whole bovine B50/GAP-43). B50/GAP-43 protein was further separated into phosphorylated and dephosphorylated species by calmodulin-Sepharose chromatography. The amounts of phosphorylated and dephosphorylated B50/GAP-43 forms were determined by electrophoresis, protein staining, and densitometry. Data on the relative phosphorylation of B50/GAP-43 protein in membrane and cytoplasmic fractions show a 10-fold difference in the ratio of cytoplasmic/membrane phosphorylation of B50/GAP-43 in AD brains with high neurofibrillary tangle (NFT) density compared to AD brains with low NFT density. This difference is due to a decreased percentage of phosphorylated B50/GAP-43 in the membrane fraction relative to that in the cytosolic fraction from high NFT density. No analogous relationship was found between the phosphorylation of B50/GAP-43 and the density of neuritic plaques in the brains examined. These data indicate differential distribution of phosphorylated and dephosphorylated B50/GAP-43 in normal and AD brains is related to NFT density but not to neuritic plaque density.