Characterization of the major human STAG3 variants using some proteomics and bioinformatics assays

• Main Authors: Inam J. Lafta, Bassam K. Kudhair, Noralhuda N. Alabid
• Format: Article
• Language: English
• Published: SpringerOpen 2020-03-01
• Series: Egyptian Journal of Medical Human Genetics
• Subjects: STAG3 gene; Variants; Immunoblotting; Immunoprecipitation; Bioinformatics
• Online Access: http://link.springer.com/article/10.1186/s43042-020-0051-0

Abstract Background STAG3 is the meiotic component of cohesin and a member of the Cancer Testis Antigen (CTA) family. This gene has been found to be overexpressed in many types of cancer, and recently, its variants have been implicated in other disorders and many human diseases. Therefore, this study aimed to analyze the major variants of STAG3. Western blot (WB) and immunoprecipitation (IP) assays were performed using two different anti-STAG3 antibodies that targeted the relevant protein in MCF-7, T-47D, MDA-MB-468, and MDA-MB-231 breast cancer cells with Jurkat and MCF-10A cells as positive and negative controls, respectively. In silico analyses were searched to study the major isoforms. Results WB and IP assays revealed two abundant polypeptides < 191 kDa and ~ 75 kDa in size. Specific bioinformatics tools successfully determined the three-dimensional (3-D) structure, the subcellular localization, and the secondary structures of the isoforms. Furthermore, some of the physicochemical properties of the STAG3 proteins were also determined. Conclusions The results of this study revealed the power of applying the biological techniques (WB and IP) with the bioinformatics assays and the possibility of their exploitation in understanding diseased genes. Exploring the major variants of STAG3 at the protein level could help decipher some disorders associated with their occurrence, along with designing drugs effective at least for some relevant diseases.