Cytochalasin D‐mediated hyperinduction of the substrate‐associated 52‐kilodalton protein p52 in rat kidney fibroblasts
Cytochalasin D‐mediated hyperinduction of the substrate‐associated 52‐kilodalton protein p52 in rat kidney fibroblasts
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细胞松弛素 D 介导的大鼠肾成纤维细胞中底物相关 52 道尔顿蛋白 p52 的过度诱导
DOI：
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发表时间：
1989
影响因子：
5.6
通讯作者：
P. Chaudhari
中科院分区：
文献类型：
--
作者：
Paul J. Hiccins;M. Ryan;P. Chaudhari
在培养的哺乳动物细胞中，某些分化和管家功能的调节受到细胞形状的显著影响。因此，使用形状调节剂细胞松弛素D（CD）来阐明影响正常大鼠肾（NRK）成纤维细胞主要52 kDa分泌/底物相关蛋白（p52）合成的潜在细胞结构影响。生物合成标记实验表明，用CD处理NRK细胞使M，52，000蛋白的培养基含量增加了10 - 18倍。二维凝胶电泳和肽片段图谱证实，CD处理后大量产生的52 kDa蛋白与NRK细胞组成型表达的p52相同。还分离出了较低分子量的蛋白质（p50; M，50，000），根据pl微异质性、蛋白酶片段化特征和对衣霉素的敏感性，可将其鉴定为p52的低糖基化形式。p50和p52在NRK和NRK/CD细胞的基质和培养基中均被检测到。然而，CD诱导和未诱导细胞的基质p52含量显著高于（200 - 500倍）相应的培养基水平。这种差异区室化、p52在NRK/CD细胞基质中积累的时间过程与其在培养基中的出现相比，以及p52从基质中脉冲追逐的动力学共同表明基质是p52沉积的初始位点。低水平的CD（1 μM）导致细胞微丝的广泛破坏，但不会导致整体细胞形状改变，也不会过度诱导p52。形态学变圆（在10-100 μM CD中观察到）与p52产生增加一致。在NRK细胞中，从扁平表型到圆形表型的转变，或者至少产生足够的微丝碎片以损害细胞基质粘附性，似乎是CD介导的p52过度诱导的一个重要方面。
Regulation of certain differentiated and housekeeping functions in cultured mammalian cells is significantly influenced by cell shape. The shape‐modulating agent cytochalasin D (CD) was used, therefore, to elucidate potential cytoarchitectural influences affecting synthesis of a major 52 kDa secreted/substrateassociated protein (p52) of normal rat kidney (NRK) fibroblasts. Biosynthetic labeling experiments indicated that treatment of NRK cells with CD increased, by 10‐18‐fold, the medium content of an M, 52,000 protein. Two‐dimensional gel electrophoresis and peptide fragment mapping confirmed that the 52 kDa protein produced in abundance as a consequence of CD treatment was identical to p52 constitutively expressed by NRK cells. A lower mw protein (p50; M, 50,000) was also resolved which, based on pl microheterogeneity, protease fragmentation profile, and sensitivity to tunicamycin, could be identified as a less‐glycosylated form of p52. p50 and p52 were both detected in the matrix and medium compartments of NRK and NRK/CD cells. The matrix p52 content of CD‐induced and uninduced cells, however, was significantly greater (by 200‐500‐fold) than the corresponding medium levels. This differential compartmentalization, the time course of p52 accumulation in the matrix of NRK/CD cells compared to its appearance in the medium, and the kinetics of p52 pulse‐chase from the matrix collectively indicated that the matrix is the initial site of p52 deposition. Low levels of CD (1 μM) produced extensive disruptions of cellular microfilaments but did not result in an overall cell shape change nor a hyperinduction of p52. Morphologic rounding (seen in 10–100 μM CD) coincided with augmented p52 production. Transition from a flat to a round phenotype in NRK cells, or at least the generation of sufficient microfilament fragmentation to compromise cellsubstrate adhesivity, appears to be an essential aspect of CD‐mediated p52 hyperinduction.
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DOI：
10.1016/s0021-9258(17)42691-3
发表时间：
1984-12
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. V. van Mourik;D. Lawrence;D. Loskutoff
通讯作者：
J. V. van Mourik;D. Lawrence;D. Loskutoff
影响因子：
2.9
作者：
SWALLA, BJ;SOLURSH, M
通讯作者：
SOLURSH, M
DOI：
--
发表时间：
1988
期刊：
The American journal of pathology
影响因子：
--
作者：
P. Phillips;P. Higgins;A. Malik;M. Tsan
通讯作者：
P. Phillips;P. Higgins;A. Malik;M. Tsan
成纤维细胞与含纤连蛋白的基质的紧密接触和局部接触粘附。
DOI：
--
发表时间：
1985
期刊：
Federation proceedings
影响因子：
--
作者：
Lark,MW;Laterra,J;Culp,LA
通讯作者：
Culp,LA
DOI：
10.1073/pnas.85.7.2161
发表时间：
1988-04-01
影响因子：
11.1
作者：
BENZEEV, A;ROBINSON, GS;FARMER, SR
通讯作者：
FARMER, SR