Fractionation and initial characterization of the kinetochore from mammalian metaphase chromosomes.
Fractionation and initial characterization of the kinetochore from mammalian metaphase chromosomes.
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DOI：
10.1083/jcb.101.3.1124
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发表时间：
1985-09
期刊：
影响因子：
--
通讯作者：
Brinkley BR
中科院分区：
文献类型：
--
作者：
Valdivia MM;Brinkley BR
我们已经从哺乳动物中期染色体中部分分离出着丝点和相关的着丝粒结构。采用硬皮病CREST（钙质沉着症、雷诺现象、食道运动障碍、硬指症、毛细血管扩张）患者自身抗体作为免疫荧光探针监测分离。程序包括用微球菌核酸酶消化总染色体DNA，用肝素去组化，并用洗涤剂和尿素解离剩余物质。我们使用密度梯度（甲硝唑胺）获得染色物质（着丝点）的富集部分。电镜下观察，着丝粒部分含有许多免疫过氧化物酶阳性的小团块，其形态与完整中期染色体的着丝粒/着丝粒区域相似。含有着丝粒成分的颗粒部分占总染色体蛋白质的5%以下，占总DNA的1%以下。CREST抗血清免疫印迹法鉴定了两个18和80 kD的多肽为着丝点抗原。在本文中，我们讨论了这些着丝粒多肽与相关着丝粒染色质的分布。
We have partially isolated the kinetochore and associated centromeric structures from mammalian metaphase chromosomes. Human autoantibodies from scleroderma CREST (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia) patients were used as immunofluorescent probes to monitor fractionation. The procedure includes digestion of total chromosomal DNA with micrococcal nuclease, dehistonization with heparin, and dissociation of the remaining material with detergent and urea. We used a density gradient (metrizamide) to obtain an enriched fraction of stained material (kinetochore). When examined by electron microscopy, the kinetochore fraction is seen to contain numerous small immunoperoxidase-positive masses which are morphologically similar to the centromere/kinetochore region of intact metaphase chromosomes. The particulate fraction that contains kinetochore components represents less than 5% of total chromosomal proteins and contains less than 1% of total DNA. Two polypeptides of 18 and 80 kD were identified as kinetochore antigens by immunoblotting with CREST antiserum. In this paper we discuss the distribution of these kinetochore polypeptides with the associated centromeric chromatin.
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DOI：
10.1083/jcb.98.1.352
发表时间：
1984-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Earnshaw WC;Halligan N;Cooke C;Rothfield N
通讯作者：
Rothfield N
DOI：
10.1083/jcb.84.1.151
发表时间：
1980-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bergen LG;Kuriyama R;Borisy GG
通讯作者：
Borisy GG
影响因子：
64.5
作者：
CAPCO, DG;WAN, KM;PENMAN, S
通讯作者：
PENMAN, S
影响因子：
3.7
作者：
GOODERHAM, K;JEPPESEN, P
通讯作者：
JEPPESEN, P
DOI：
10.1073/pnas.77.3.1627
发表时间：
1980-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
MOROI, Y;PEEBLES, C;TAN, EM
通讯作者：
TAN, EM