非肌肉细胞经曲拉通提取的细胞骨架组分中的腺嘌呤和鸟嘌呤核苷酸含量。

Adenine and guanine nucleotide content of Triton-extracted cytoskeletal fractions of nonmuscle cells.

作者信息

Angelastro J M, Purich D L

机构信息

Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville 32610-0245.

出版信息

Anal Biochem. 1992 Jul;204(1):47-52. doi: 10.1016/0003-2697(92)90137-v.

DOI:10.1016/0003-2697(92)90137-v

PMID:1514695

Abstract

Determination of the adenine and guanine nucleotides in Triton X-100-extracted cytoskeletal fractions was utilized to estimate the actin and tubulin content of the assembled cytoskeletons in nonmuscle cells. Results with stable cell lines (i.e., rat pheochromocytoma PC12 and neuroblastoma NB41A3) and with primary cultures (i.e., human foreskin fibroblasts and chick embryonic dorsal root ganglion neurons) exhibited levels of cytoskeletal fraction ADP and GDP consistent with their assembly-induced nucleoside-5'-triphosphatase activities only previously analyzed in vitro. Likewise, estimates of actin and tubulin content fall in the range of values obtained by other experimental approaches. In contrast, analysis of whole cell nucleotides showed high [ATP]/[ADP] and [GTP]/[GDP] ratios, suggesting there is little, if any, contamination of the cytoskeletal nucleotide pool by other cellular nucleotides.

摘要

利用对Triton X-100提取的细胞骨架组分中腺嘌呤和鸟嘌呤核苷酸的测定，来估计非肌肉细胞中组装好的细胞骨架的肌动蛋白和微管蛋白含量。稳定细胞系（即大鼠嗜铬细胞瘤PC12和神经母细胞瘤NB41A3）以及原代培养物（即人包皮成纤维细胞和鸡胚背根神经节神经元）的结果显示，细胞骨架组分中ADP和GDP的水平与之前仅在体外分析过的其组装诱导的核苷-5'-三磷酸酶活性一致。同样，肌动蛋白和微管蛋白含量的估计值落在通过其他实验方法获得的值的范围内。相比之下，对全细胞核苷酸的分析显示出高的[ATP]/[ADP]和[GTP]/[GDP]比率，这表明细胞骨架核苷酸库几乎没有受到其他细胞核苷酸的污染（如果有污染的话也极少）。