Sw. Rothwell et al., AFFINITY PURIFICATION AND SUBCELLULAR-LOCALIZATION OF KINESIN IN HUMAN NEUTROPHILS, Journal of leukocyte biology, 53(4), 1993, pp. 372-380
Studies of granule-microtubule interactions in human neutrophils have
suggested that mechanochemical ATPases such as kinesin or dynein may p
lay a role in granule mobilization during neutrophil activation by inf
lammatory signals. In this study we show that proteins extracted from
the surface of neutrophil granules, found previously to contain microt
ubule-dependent ATPase activity, caused microtubules polymerized from
phosphocellulose-purified rat brain tubulin to move across glass slide
s. Antibodies were generated against peptides based on two regions of
the amino acid sequence of Drosophila kinesin: the ATPase active site
(amino acids 86-99) in the head of the kinesin heavy chain and the tai
l of the heavy chain (residues 913-933). These antibodies were found t
o recognize kinesin in rat brain extracts as well as kinesin-like poly
peptides in extracts of human neutrophils. Furthermore, when used in i
mmunoaffinity chromatography, these antibodies permitted the isolation
of a protein from neutrophil granule extracts that was recognized by
Drosophila kinesin antibodies. Subcellular localization by immunofluor
escence microscopy showed this protein to be associated principally wi
th the cytoplasmic granules of neutrophils.