FGF-FGFR signaling mediated through glycosaminoglycans in microtiter plate and cell-based microarray platforms.
FGF-FGFR signaling mediated through glycosaminoglycans in microtiter plate and cell-based microarray platforms.
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DOI：
10.1021/bi401284r
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发表时间：
2013-12-17
期刊：
影响因子：
2.9
通讯作者：
Linhardt, Robert J.
中科院分区：
文献类型：
--
作者：
Sterner, Eric;Meli, Luciana;Kwon, Seok-Joon;Dordick, Jonathan S.;Linhardt, Robert J.
成纤维细胞生长因子（FGF）通过其与成纤维细胞生长因子受体（FGFR）和糖胺聚糖（GAG）共受体的相互作用而发出细胞生长信号。在这里，我们研究了五种不同的FGF（FGF 1，FGF 2，FGF 6，FGF 8和FGF 8b）通过FGFR 3c的信号传导。筛选GAG和GAG衍生物共受体的小文库以更好地理解这些共受体对信号传导的结构-活性关系。最初，在基于微量滴定孔的细胞增殖测定中收集数据。为了减少试剂需求并提高测定通量，开发了基于细胞的微阵列平台。在该基于细胞的微阵列中，将表达FGFR 3c的细胞印刷在约30 nL的藻酸盐水凝胶液滴中，并与FGF和GAG一起孵育。肝素是所有研究的FGF的最有效的GAG共受体。其他GAG，如2-O-乙酰肝素和硫酸软骨素B，也是有效的共受体。FGF 8和FGF 8b的信号传导显示出对共受体结构的最宽耐受性。最后，这种芯片上基于细胞的微阵列提供了与基于微量滴定孔的测定相当的数据，表明共受体测定可以转化为高通量测定。
Fibroblast growth factor (FGF) signals cell growth through its interaction with a fibroblast growth factor receptor (FGFR) and a glycosaminoglycn (GAG) co-receptor. Here we examine the signaling of five different FGFs (FGF1, FGF2, FGF6, FGF8 and FGF8b) through FGFR3c. A small library of GAG and GAG-derivative co-receptors are screened to better understand the structure-activity relationship of these co-receptors on signaling. Initially, data were collected in a microtiter well-based cell proliferation assay. In an effort to reduce reagent requirements and improve assay throughput a cell-based microarray platform was developed. In this cell-based microarray, FGFR3c expressing cells were printed in alginate hydrogel droplets of ~30 nL and incubated with FGF and GAG. Heparin was the most effective GAG co-receptor for all FGFs studied. Other GAGs, such as 2-O-desulfated heparin and chondroitin sulfate B, were also effective co-receptors. Signaling by FGF8 and FGF8b showed the widest tolerance for co-receptor structure. Finally, this on-chip cell-based microarray provides comparable data to a microtiter well-based assay, demonstrating that the co-receptor assay can be converted into a high throughput assay.
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影响因子：
3.8
作者：
Fernandes TG;Kwon SJ;Bale SS;Lee MY;Diogo MM;Clark DS;Cabral JM;Dordick JS
通讯作者：
Dordick JS
DOI：
10.1016/j.bbagen.2008.09.001
发表时间：
2009-01-01
影响因子：
3
作者：
Asada, Masahiro;Shinomiya, Michiyo;Imamura, Toru
通讯作者：
Imamura, Toru
DOI：
10.1073/pnas.81.4.1030
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ATHA, DH;STEPHENS, AW;ROSENBERG, RD
通讯作者：
ROSENBERG, RD
影响因子：
56.9
作者：
ORNITZ, DM;HERR, AB;WAKSMAN, G
通讯作者：
WAKSMAN, G
从肝素污染中学到的经验教训。
影响因子：
11.9
作者：
Liu H;Zhang Z;Linhardt RJ
通讯作者：
Linhardt RJ