Purification of kinesin-related protein complexes from eggs and embryos

D. G. Cole, J. M. Scholey, S. Endow, C. Schutt, E. Young, D. Hackney, D. White, S. Block, P. Satir

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

We have developed a biochemical screen for the identification of kinesin- related proteins (KRPs) in their natural host cells and the subsequent purification of these KRPS as native, functional multimeric complexes. The screen involves immunoblotting with pan-kinesin peptide antibodies that recognize several presumptive KRPs in cytosolic extracts; the antibodies have been used so far to monitor the purification of two bona fide kinesin- related motor protein complexes. These two KRPs were purified via AMPPNP- induced microtubule affinity binding, ATP-induced elution from AMPPNP microtubules, gel filtration fractionation, and sucrose density gradient centrifugation. KRP(85/95) from sea urchin (Strongylocentrotus purpuratus) eggs behaves as a heterotrimeric complex of 85-, 95-, and 115- kDa subunits that moves toward the plus ends of microtubule tracks at approximately 0.4 μm/s. KRP(130) from fruitfly (Drosophila melanogaster) embryos behaves as a homotetrameric complex of four 130-kDa subunits that moves toward the plus ends of microtubule tracks at approximately 0.04 μm/s. To our knowledge, KRP(85/95) and KRP(130) are the only KRPS to have been purified from native tissue as functional multimeric motor complexes.

Original languageEnglish (US)
Pages (from-to)158s-162s
JournalBiophysical journal
Volume68
Issue number4 SUPPL.
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Biophysics

Fingerprint Dive into the research topics of 'Purification of kinesin-related protein complexes from eggs and embryos'. Together they form a unique fingerprint.

  • Cite this

    Cole, D. G., Scholey, J. M., Endow, S., Schutt, C., Young, E., Hackney, D., White, D., Block, S., & Satir, P. (1995). Purification of kinesin-related protein complexes from eggs and embryos. Biophysical journal, 68(4 SUPPL.), 158s-162s.