Isolation of mitotic microtubule-associated proteins from sea urchin eggs.

Abstract

We have used a taxol-based microtubule purification procedure and monoclonal antibodies to isolate and characterize the MAPs of mitotic spindle microtubules in the fertilized sea urchin egg. In so doing, we hope to have identified some of the essential working parts of the mitotic apparatus, namely those proteins that regulate the assembly, disassembly, organization and mechanochemical properties of spindle microtubules. The results of this effort strongly suggest that a rich diversity of polypeptides associate with mitotic spindle microtubules. Whether each of these represents an individual protein species is not currently known. It is possible, for example, that particular spindle MAPs comprise multiple, distinct subunits. This would not be surprising in light of the facts that both MAP-1 and MAP-2 contain lower molecular weight subunits, and that axonemal dyneins are complex assemblies of several polypeptide species. Our future efforts with the sea urchin system will be to determine how the various mitotic spindle MAPs we have identified function individually and in concert, and how those functions contribute to the mechanochemical properties of the spindle.