Differential effects of cobalt on the initiation of fast axonal transport

Effects of Co2+ on the fast axonal transport of individual proteins were examined in vitro in bullfrog spinal/sciatic nerves. 35S-methionine-labeled proteins, fast-transported in control and Co2+-treated preparations were separated via two-dimensional gel electrophoresis. While the overall amount of protein transported was reduced, no qualitative differences could be seen when gel fluorographic patterns were compared. Quantitative analyses of the 48 most abundantly transported species revealed two significantly different populations (p less than 0.01) differentially sensitive to Co2+ and distinguishable to a large extent by molecular weight. Those proteins less sensitive to Co2+ ranged from approximately 20,000 to 35,000 daltons while those more sensitive to Co2+ were greater than approximately 35,000 daltons. The finding that all proteins are affected by Co2+ supports the proposal that fast-transported proteins are subject to a common Co2+-sensitive, Ca2+-requiring step. The observed differential effects are consistent with more than one Ca2+-dependent step occurring during the initiation phase of fast transport.