Revisiting the mystery of fibronectin multimers: the fibronectin matrix is composed of fibronectin dimers cross-linked by non-covalent bonds.
Revisiting the mystery of fibronectin multimers: the fibronectin matrix is composed of fibronectin dimers cross-linked by non-covalent bonds.
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重温纤连蛋白多聚体之谜：纤连蛋白基质是由非共价键交联的纤连蛋白二聚体组成。
DOI：
10.1016/j.matbio.2009.03.002
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Erickson,HaroldP
中科院分区：
文献类型：
--
作者：
Ohashi,Tomoo;Erickson,HaroldP
纤维连接蛋白(FN)基质纤维一直被认为是由二硫键结合的FN多聚体形成的，尽管没有直接证据表明它们之间是共价连接的。为了了解这些纤维的生化性质，我们用0.2%的脱氧胆酸盐和DNA酶从FN-YPet转染的3T3细胞培养中提取了粗制的FN基质。不溶性提取的基质保留了纤维状结构，大部分提取的蛋白质在还原条件下以FN单体的形式在十二烷基硫酸钠凝胶上迁移。在非还原条件下，一些FN分子似乎被捕获在堆积凝胶的顶部。我们在将荧光标记的FN二聚体与提取的基质混合后加载到十二烷基硫酸钠凝胶中进行了测试，发现它们中的大多数与提取的蛋白质一起被捕获在堆积凝胶的顶部。这些结果表明，提取的基质中的某些组分堵塞了堆积凝胶，FN二聚体被捕获。旋转阴影电子显微镜显示，提取的基质中有一些类似于纤维蛋白微纤维的纤维。多肽质量指纹图谱证实提取的基质中存在纤维蛋白。已知纤维蛋白可以形成二硫键多聚体，它很可能是该体系中堵塞堆积凝胶和捕获FN分子的成分之一。因此，FN分子作为多聚体在十二烷基硫酸钠凝胶上迁移的现象是由于提取物中其他大组分的存在而产生的假象。我们得出结论，FN基质纤维是由FN二聚体通过非共价蛋白质-蛋白质键进一步交联而成的。
Fibronectin (FN) matrix fibrils have long been thought to be formed by disulfide-bonded FN multimers, although there is no direct evidence that they are covalently linked with each other. To understand the biochemical properties of these fibrils, we extracted a crude FN matrix from FN-YPet transfected 3T3 cell culture using 0.2% deoxycholate and DNase. The insoluble extracted matrix preserved fibrillar structures and a major portion of the extracted proteins migrated as FN monomers on an SDS gel under reducing conditions. Under non-reducing conditions, some FN molecules appeared to be trapped at the top of the stacking gel. We tested this by mixing fluorescently labeled FN dimers with the extracted matrix just before loading on an SDS gel, and found that most of them were trapped with the extracted proteins at the top of the stacking gel. These results suggested that some components of the extracted matrix plugged the stacking gel and FN dimers were trapped with them. Rotary shadowing electron microscopy showed that the extracted matrix had some fibers that resembled fibrillin microfibrils. Peptide mass fingerprinting confirmed the presence of fibrillin in the extracted matrix. Fibrillin is known to form disulfide-bonded multimers and it is likely to be one of the components that plug the stacking gel and trap FN molecules in this system. The phenomenon by which FN molecules appear to migrate as multimers on SDS gels is thus an artifact rising from the presence of other large components in the extract. We conclude that FN matrix fibrils are made of FN dimers that are further cross-linked by non-covalent protein–protein bonds.
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DOI：
10.1016/0006-291x(77)90359-x
发表时间：
1977-01-01
影响因子：
3.1
作者：
KESKIOJA, J;MOSHER, DF;VAHERI, A
通讯作者：
VAHERI, A
纤维蛋白原的三结结构：通过阴影和负染色电子显微镜确认
影响因子：
6.7
作者：
W. Fowler;Harold P. Erickson
通讯作者：
Harold P. Erickson
通过重组纤连蛋白的过度表达来抑制人纤维肉瘤细胞的转化表型。
影响因子：
11.2
作者：
Hiroki Akamatsu;Keiko Ichihara;Keiichi Ozono;Wataru Kamiike;Hikaru Matsuda;Kiyotoshi Sekiguchi
通讯作者：
Kiyotoshi Sekiguchi
影响因子：
3.3
作者：
Sabatier, Laetitia;Chen, Daliang;Reinhardt, Dieter P.
通讯作者：
Reinhardt, Dieter P.
DOI：
10.1083/jcb.124.5.855
发表时间：
1994-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Zhang H;Apfelroth SD;Hu W;Davis EC;Sanguineti C;Bonadio J;Mecham RP;Ramirez F
通讯作者：
Ramirez F