Hu Yanhui, Gao Chenxi, Mckenna William, Xia Baolong, Ariss Majd, Mohr Stephanie, Perrimon Norbert
Department of Genetics, Blavatnik Institute, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Cell Signaling Technology, 3 Trask Lane, Danvers, MA 01923, USA.
bioRxiv. 2024 Dec 22:2024.12.20.629730. doi: 10.1101/2024.12.20.629730.
Signaling pathways play key roles in many important biological processes such as cell division, differentiation, and migration. Phosphorylation site-specific antibodies specifically target proteins phosphorylated on a given tyrosine, threonine, or serine residue. Use of phospho-specific antibodies facilitates analysis of signaling pathway regulation and activity. Given the usefulness of phospho-specific antibodies, a number of collections of these antibodies have been generated, typically for detection of phosphorylated mammalian proteins. Anecdotal evidence shows that some of these are also useful for detection of phosphorylated forms of orthologous proteins in model organisms. We propose that anti-phospho-mammalian protein antibody collections comprise an untapped resource for research in other species. To systematically analyze the potential utility of anti-phospho-mammalian protein antibodies in other species, we developed the Cross-species Epitope Sequence Analysis software tool (CESA). CESA identifies and aligns orthologous proteins in model species, then analyzes conservation of antibody target sites. We used CESA to predict what phospho-specific antibodies in a collection from Cell Signaling Technology (CST) might be useful for studies in and other species. CESA predicts that more than 232 sites on 116 proteins can potentially be targeted by the antibodies initially developed at CST to detect human, mouse, or rat phosphoproteins.
信号通路在许多重要的生物学过程中发挥着关键作用,如细胞分裂、分化和迁移。磷酸化位点特异性抗体专门针对在特定酪氨酸、苏氨酸或丝氨酸残基上磷酸化的蛋白质。使用磷酸化特异性抗体有助于分析信号通路的调节和活性。鉴于磷酸化特异性抗体的实用性,已经产生了许多这类抗体的集合,通常用于检测磷酸化的哺乳动物蛋白质。轶事证据表明,其中一些抗体也可用于检测模式生物中直系同源蛋白质的磷酸化形式。我们提出,抗磷酸化哺乳动物蛋白抗体集合是其他物种研究中尚未开发的资源。为了系统地分析抗磷酸化哺乳动物蛋白抗体在其他物种中的潜在效用,我们开发了跨物种表位序列分析软件工具(CESA)。CESA可识别并比对模式物种中的直系同源蛋白质,然后分析抗体靶位点的保守性。我们使用CESA来预测细胞信号技术公司(CST)的一个抗体集合中的哪些磷酸化特异性抗体可能对[具体物种]和其他物种的研究有用。CESA预测,最初由CST开发用于检测人、小鼠或大鼠磷酸化蛋白质的抗体可能靶向116种蛋白质上的232多个位点。