The carboxyl-terminal domain of kinesin heavy chain is important for membrane binding
Sea urchin kinesin is a plus end-directed microtubule-based motor consisting of two heavy chains and two light chains and is proposed to be responsible (a) for the transport of membranous organelles along microtubules in sea urchin mitotic spindles (Wright, B. D., Henson, J. H., Wedaman, K. P., Will...
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Published in | The Journal of biological chemistry Vol. 269; no. 2; pp. 1477 - 1485 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Bethesda, MD
American Society for Biochemistry and Molecular Biology
14.01.1994
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Subjects | |
Online Access | Get full text |
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Summary: | Sea urchin kinesin is a plus end-directed microtubule-based motor consisting of two heavy chains and two light chains and
is proposed to be responsible (a) for the transport of membranous organelles along microtubules in sea urchin mitotic spindles
(Wright, B. D., Henson, J. H., Wedaman, K. P., Willy, P. J., Morand, J. N., and Scholey, J. M. (1991) J. Cell Biol. 113, 817-833)
and (b) for the radial dispersion of endoplasmic reticulum and endosomal membranes in non-mitotic cultured coelomocytes (Henson,
J. H., Nesbitt, D., Wright, B. D., and Scholey, J. M. (1992) J. Cell Sci. 103, 309-320). We report here that sea urchin kinesin
is indeed able to bind in a concentration-dependent and saturable manner to microsomal membranes isolated from sea urchin
eggs in the presence of MgATP. The kinesin light chains may not be essential for membrane binding since kinesin containing
negligible amounts of light chains binds as well as kinesin containing stoichiometric amounts of light chains. Finally, we
propose that kinesin binds to membranes with the carboxyl-terminal domain of the heavy chain (amino acid residues 858-1031)
since the bacterially expressed and then isolated stalk-tail fragment of kinesin heavy chain, in contrast to the stalk fragment,
is able (a) to bind membranes in a concentration-dependent and saturable manner and (b) to compete with native kinesin for
membrane binding. Our results support the hypothesis that the carboxyl-terminal domains of the heavy chains attach kinesin
molecules to their membranous cargo in mitotic and interphase sea urchin cells. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0021-9258 1083-351X |
DOI: | 10.1016/s0021-9258(17)42281-2 |