Promotion of microtubule assembly in vitro by a novel 35-kDa protein purified from human term placenta.
Microtubule assembly promoting-protein (taxol-like protein, TALP) was purified by combination of high salt extraction, phosphocellulose chromatography and hydroxyapatite chromatography from human term placenta. Molecular weight of purified protein was identified as 35kDa on SDS-polyacrylamide gel electrophoresis. In vitro, TALP promoted microtubule assembly in dose-dependent manner in spite of the absence of GTP. Both TALP (0.5 microM) and taxol (10 microM) gave hyperbolic kinetics and shortened the lag time for microtubule polymerization. TALP-stabilized microtubules were resistant to depolymerization by cold (4 degrees C) and CaCl2 (4 mM) like taxol-stabilized microtubules. TALP showed its direct binding on the microtubule in cosedimentation assay. These results suggest that the action mechanism of TALP on the microtubule assembly is similar to taxol in vitro and the binding site of TALP is available on the intact microtubule.
Hwang
B D
BD
Department of Biochemistry, College of Medicine Chungnam National University, Daejeon, Korea.
Kwak
S T
ST
Kweon
G R
GR
Lim
K
K
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007700:Kinetics; D008868:Microtubule Proteins / Q000187:drug effects / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects* / Q000648:ultrastructure; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D013997:Time Factors; D014404:Tubulin / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
D008868:Microtubule Proteins; D014404:Tubulin