Cande W Z, Wolniak S M
J Cell Biol. 1978 Nov;79(2 Pt 1):573-80. doi: 10.1083/jcb.79.2.573.
Mitotic PtK1 cells, lysed at anaphase into a carbowax 20 M Brij 58 solution, continue to move chromosomes toward the spindle poles and to move the spindle poles apart at 50% in vivo rates for 10 min. Chromosome movements can be blocked by adding metabolic inhibitors to the lysis medium and inhibition of movement can be reversed by adding ATP to the medium. Vanadate at micromolar levels reversibly inhibits dynein ATPase activity and movement of demembranated flagella and cilia. It does not affect glycerinated myofibril contraction or myosin ATPase activty at less than millimolar concentrations. Vanadate at 10--100 micron reversibly inhibits anaphase movement of chromosomes and spindle elongation. After lysis in vanadate, spindles lose their fusiform appearance and become more barrel shaped. In vitro microtubule polymerization is insensitive to vanadate.
处于有丝分裂后期的PtK1细胞,在被裂解于聚乙二醇20M Brij 58溶液中后,会以体内速率的50%继续将染色体移向纺锤体极,并使纺锤体极分开10分钟。通过向裂解培养基中添加代谢抑制剂可阻断染色体运动,而向培养基中添加ATP可逆转运动抑制。微摩尔浓度的钒酸盐可可逆地抑制动力蛋白ATP酶活性以及去膜鞭毛和纤毛的运动。在小于毫摩尔浓度时,它不影响甘油化肌原纤维收缩或肌球蛋白ATP酶活性。10 - 100微摩尔的钒酸盐可可逆地抑制染色体的后期运动和纺锤体伸长。在钒酸盐中裂解后,纺锤体失去其纺锤形外观,变得更呈桶状。体外微管聚合对钒酸盐不敏感。