These different ‘species’ of Cys can be “labeled” with charges that will then separate out the reduced Cys from the oxidized Cys residues using the PAGE method. These thiols can be modified in a number of different ways, but this article will focus on whether the thiol is reduced (SH), or oxidized and exists in a disulfide bond (S-S) with another Cys (Figure 1). PEMSA is a useful method to determine whether a certain condition is affecting the redox state of your proteins and therefore if the Cys residues are reduced or oxidized (Figure 1).
#Pemsa carreers free
A protein with reduced Cys have free thiols, whereas the oxidized form may engage in a disulfide bond with another Cys in the same protein. Reduced and oxidized Cys residues in a protein. There are even antibodies that can detect various phosphorylated proteins, but detecting redox modified proteins is a bit more involved as discussed below.
Just like phosphorylation signaling cascades, which is a more common post-translational modification, redox modifications can cause activation/deactivation of pathways.
If you want to be able to determine redox modifications to the cysteine (Cys) residues found in your protein of interest, or if certain conditions alter the protein redox state, then PEMSA might work for you! For example, you may want to know why your transcription factor is not binding to a DNA promoter, or why you cannot activate a certain pathway. This method is essentially an agarose gel electrophoresis technique that detects protein:nucleic acid interactions, as the mobility of the labeled nucleic acid will be retarded if bound to a protein (compared to unbound DNA).Ī lesser-known technique is the PEMSA, or protein electrophoretic mobility shift assay, which uses the polyacrylamide gel electrophoresis (PAGE) method and is more like a redox (reduction-oxidation reaction) western blot. I set myself the goal of slowly building my own business and getting exposure to as many different industries as possible.Studying nucleic acid interactions with proteins can be accomplished using a rapid and efficient electrophoretic mobility shift assay (EMSA). At that time, I had started freelancing for family and friends who were in need of legal advice and when the lockdown hit, I realised that I needed to step up my game. However, I soon realised that I did not enjoy working in a corporate environment and wanted to focus on developing myself, performing impactful work and building interpersonal relationships. One thing led to another and I ended up working as DP Lead for a tech giant. This has been my most formative experience to date and it is also where I discovered Data Privacy, right when the GDPR hype was starting. Here, I had first-hand experience with diverse clients on commercial, IP and data privacy legal matters. When I graduated from university, I rejected a position in a global business and opted instead for a much more meaningful role in a small legal-tech startup. This has led to some interesting experiences that I have found personally and professionally enriching - including working with Flex. Looking back, I’ve always been a genuinely curious person and have strived to carve my own path rather than just listen to friends and family.
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