Identification of immunoreactive regions of homology between soluble epidermal growth factor receptor and α5-integrin.
Identification of immunoreactive regions of homology between soluble epidermal growth factor receptor and α5-integrin.
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DOI：
10.1021/bi200126j
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发表时间：
2011-05-24
期刊：
影响因子：
2.9
通讯作者：
Maihle NJ
中科院分区：
文献类型：
--
作者：
Wilken JA;Baron AT;Foty RA;McCormick DJ;Maihle NJ
表皮生长因子受体（EGFR/HER1/ERBB1）基因编码的蛋白正被研究作为许多人类癌症的诊断、预后和治疗性生物标志物。这些组织/肿瘤生物标志物的临床应用受到限制，部分原因是使用不同的免疫试剂观察到的EGFR表达结果不一致。先前的研究使用了EGFR定向抗体，不能区分全长EGFR和可溶性EGFR （sEGFR）的表达。我们已经生成并鉴定了一种针对位于sEGFR独特的78个氨基酸羧基末端序列中的31个肽段（残基604-634）的抗sEGFR多克隆抗血清。在这里，我们使用这种抗体来证明sEGFR在许多癌源性细胞系中与EGFR共表达。此外，我们发现该抗体还能检测到~140-kDa （p140）的第二种蛋白。严格的生化鉴定鉴定出第二种蛋白为α5整合素。研究人员发现，α5-整合素小腿区域的26个氨基酸肽（残基710-35）与31个羧基末端的sEGFR肽具有35%的序列同源性，并且对抗sEGFR的亲和力比同源的31个片段的sEGFR肽低约5倍。我们得出结论，sEGFR的羧基末端和α5-整合素的calf-1结构域共享一个序列相同的区域，这导致它们与抗sEGFR相互免疫反应。我们还证明了抗sEGFR在体外促进3D组织内聚和压实，进一步表明sEGFR和α5整合素之间的功能联系以及calf-1结构域在细胞粘附中的作用。这些结果对EGFR和sEGFR作为癌症生物标志物的研究具有重要意义，也为细胞表面EGFR亚型和整合素在细胞粘附和生存信号传导等复杂过程中的相互作用机制提供了新的见解。
Proteins encoded by the epidermal growth factor receptor (EGFR/HER1/ERBB1) gene are being studied as diagnostic, prognostic, and theragnostic biomarkers for numerous human cancers. The clinical application of these tissue/tumor biomarkers has been limited, in part, by discordant results observed for EGFR expression using different immunological reagents. Previous studies have used EGFR-directed antibodies that cannot distinguish between full-length vs. soluble EGFR (sEGFR) expression. We have generated and characterized an anti-sEGFR polyclonal antiserum directed against a 31-mer peptide (residues 604–634) located within the unique 78 amino acid carboxy-terminal sequence of sEGFR. Here, we use this antibody to demonstrate that sEGFR is co-expressed with EGFR in a number of carcinoma-derived cell lines. In addition, we show that a second protein of ~140-kDa (p140) also is detected by this antibody. Rigorous biochemical characterization identifies this second protein to be α5-integrin. We show that a 26 amino acid peptide in the calf domain of α5-integrin (residues 710–35) shares 35% sequence identity with a 31-mer carboxy-terminal sEGFR peptide, and exhibits approximately 5 times lower affinity for anti-sEGFR than does the homologous 31-mer sEGFR peptide. We conclude that the carboxy-terminus of sEGFR and the calf-1 domain of α5-integrin share a region of sequence identity, which results in their mutual immunological reactivity with anti-sEGFR. We also demonstrate that anti-sEGFR promotes 3D tissue cohesion and compaction in vitro, further suggesting a functional link between sEGFR and α5-integrin and a role of the calf-1 domain in cell adhesion. These results have implications for the study of both EGFR and sEGFR as cancer biomarkers, and also provide new insight into the mechanisms of interaction between cell surface EGFR isoforms and integrin in complex processes such as cell adhesion and survival signaling.
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影响因子：
--
作者：
Flynn JF;Wong C;Wu JM
通讯作者：
Wu JM
影响因子：
4
作者：
Robinson, EE;Zazzali, KM;Foty, RA
通讯作者：
Foty, RA
DOI：
10.1089/153685902760173908
发表时间：
2002-06-01
期刊：
HYBRIDOMA AND HYBRIDOMICS
影响因子：
--
作者：
Christensen, TA;Reiter, JL;Maihle, NJ
通讯作者：
Maihle, NJ
影响因子：
11.2
作者：
Lafky, JM;Baron, LT;Maihle, NJ
通讯作者：
Maihle, NJ
影响因子：
11.2
作者：
Chen, Nanyue;Ye, Xiang-Cang;Lin, Sue-Hwa
通讯作者：
Lin, Sue-Hwa