Novel heterotrimeric kinesin-related protein purified from sea urchin eggs

@article{Cole1993NovelHK,
  title={Novel heterotrimeric kinesin-related protein purified from sea urchin eggs},
  author={Douglas G. Cole and Steven Chinn and Karen P. Wedaman and Kirsten Hall and T. Minh Vuong and Jonathan M. Scholey},
  journal={Nature},
  year={1993},
  volume={366},
  pages={268-270}
}
KINESIN heavy chain and kinesin-related polypeptides (KRPs) comprise a family of motor proteins with diverse intracellular transport functions1–7. Using pan-kinesin peptide antibodies that react with these proteins8,9, we haven previously purified from sea urchin eggs a trimeric microtubule-binding and bundling protein, KRP(85/95) (ref. 8) comprising subunits of Mr115,000 (115K), 95K and 85K. We report here that kinesin-related genes encode the 85K and 95K subunits, and that the protein can be… 
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273 Citations
Highly Influential Citations
21
Background Citations
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Methods Citations
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273 Citations

Purification of kinesin-related protein complexes from eggs and embryos.

KRP(85/95) and KRP(130) are the only KRPs to have been purified from native tissue as functional multimeric motor complexes.

Purification of novel kinesins from embryonic systems.

Kinesin-II, a membrane traffic motor in axons, axonemes, and spindles

This comparison shows that kinesin-II has a similar overall structural organization to kinesIn, having two motor domains lying at one end of the molecule linked by a rod to a tail where the accessory subunits are located, and appears to allow both kinesins to serve as axonal vesicle transport motors that can be deployed as membrane traffic motors in other subcellular locations such as axonemes and spindles.

KLP38B: A Mitotic Kinesin-related Protein That Binds PP1

It is demonstrated that recombinant KLP38B and PP1 associate in vitro, the first demonstration of direct binding of a kinesin-related protein to a regulatory enzyme.

The Kar3p kinesin-related protein forms a novel heterodimeric structure with its associated protein Cik1p.

Mapping of the interaction domains of the two proteins by two-hybrid analyses indicates that Kar3p and Cik1p associate in a highly specific manner along the lengths of their respective coiled-coil domains, demonstrating that the Kar3-Cik1 complex has a novel heterodimeric structure not observed previously for kinesin complexes.

Monomeric and dimeric states exhibited by the kinesin-related motor protein KIF1A.

It is predicted that native KIF1A can exist as both an inactive monomer and an active homodimer formed in part through its neck coiled-coil domain.

Purification and Characterization of Native Conventional Kinesin, HSET, and CENP-E from Mitotic HeLa Cells*

It is found that the 120-kDa HeLa cell conventional kinesin is an active motor that induces microtubule gliding at ∼30 μm/min at room temperature and does not contain light chains, although light chains were detected in other fractions.

A Molecular Motor for Microtubule-Dependent Organelle Transport in Neurospora crassa

The general belief is that variability within the tail domains reflects different functions or binding affinities of the respective motors, and is indeed supported by experimental evidence.

Heterodimerization of the two motor subunits of the heterotrimeric kinesin, KRP85/95.

It is reported that SpKRP85-SpKRP95 complexes can be immunoprecipitated from a cell-free translation system, providing direct evidence that Sp kinesin-related polypeptides are capable of heterodimerization.

Fractionation and Characterization of Kinesin II Species in Vertebrate Brain

This work isolated nucleotide‐dependent, microtubule‐binding proteins from 13‐day chick embryo brain and identified three KIF species, the most abundant species, kinesin I, which exhibited the expected long range micro tubule gliding activity, and KIF1C, which did not move microtubules.
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References

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Isolation of a sea urchin egg kinesin-related protein using peptide antibodies.

A triplet of polypeptides, which the authors refer to as KRP(85/95), binds to purified sea urchin egg tubulin in an AMPPNP-enhanced, ATP-sensitive manner and induces the formation of microtubule bundles, and it is proposed that the triplet corresponds to a novel sea urchesin egg kinesin-related protein.

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It is reported that, unlike antibodies to conventional kinesin, the peptide antibodies to the kines in motor domain immunofluorescently label spindles and kinetochores in mitotic tissue culture cells, suggesting that kinesIn-like proteins may have important roles in chromosome movement and mitosis.

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The finding that sea urchin kinesin contains equimolar quantities of heavy and light chains, together with the aforementioned data on kinesIn morphology, suggests that native sea urchesin is a tetramer assembled from two light chains and two heavy chains.

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Identification and partial characterization of six members of the kinesin superfamily in Drosophila.

The identified and partially characterized six additional members of the Drosophila kinesin superfamily that encode proteins that are 40-60% identical to the motor domain of the kinesIn heavy-chain sequence.

Isolation of a 45-kDa fragment from the kinesin heavy chain with enhanced ATPase and microtubule-binding activities.

This study uses limited proteolysis to study the structure of bovine brain kinesin, a heterotetramer composed of two heavy and two light chains, and proposes that cleavage of the heavy chain uncouples ATPase activity from its translocator activity, which may require other parts of the molecule.