1 CnaB2 spontaneously forms an intramolecular isopeptide bond between Lys31 and Asp117 ( Fig. Fundamentals of SpyTag/Sp圜atcher technology 1.1 Spontaneous amidation: kinetic and thermodynamic features SpyTag/Sp圜atcher is a protein coupling approach created by splitting the CnaB2 domain from the fibronectin binding protein FbaB from Streptococcus pyogenes. We describe limitations as well as future directions. The simple genetic encoding and rapid irreversible reaction provide diverse opportunities to enhance protein function. In living cells, SpyTag allowed imaging of protein trafficking, retargeting of CAR-T cell killing, investigation of heart contraction, and control of nucleosome position. Combinatorial increase in functionality has been achieved through modular derivatisation of antibodies, light-emitting diodes or viral vectors. For enzymes, Spy technology has increased resilience, promoted substrate channelling, and assembled hydrogels for continuous flow biocatalysis. Icosahedral multimers are being evaluated for vaccination against malaria, HIV and cancer. A toolbox of scaffolds allows SpyTag-fusions to be assembled into defined multimers, from dimers to 180-mers, or unlimited 1D, 2D or 3D networks. SpyTag/Sp圜atcher is mechanically stable, so is widely used for investigating protein folding and force sensitivity. Anchoring of SpyTag-linked proteins has been established to diverse nanoparticles or surfaces, including gold, graphene and the air/water interface. New variants of this pair allow reaction at a rate approaching the diffusion limit, while reversible versions allow purification of SpyTagged proteins or tuned dynamic interaction inside cells. ![]() ![]() SpyTag forms a spontaneous and irreversible isopeptide bond upon binding its protein partner Sp圜atcher, where both parts are genetically-encoded. ![]() Here we review SpyTag technology, now used in hundreds of publications or patents, and its potential for detecting and controlling protein behaviour. Therefore generic approaches are needed to overcome this diversity and stream-line protein analysis or application. Mod posts Serious posts Megathread Breaking news Unfilter Please use spoiler tags to hide spoilers.Proteins span an extraordinary range of shapes, sizes and functionalities. Tags to use: Use a post tag to designate your post as a serious, on-topic-only thread. If you think your post has disappeared, see spam or an inappropriate post, please do not hesitate to contact the mods, we're happy to help. Spam, machine-generated content, and karma farming are not permitted. more >ĭo not post harmful misinformation. more >Ĭomment replies consisting solely of images will be removed. Mods reserve the right to remove content or restrict users' posting privileges as necessary if it is deemed detrimental to the subreddit or to the experience of others. Soliciting money, goods, services, or favours is not allowed. ![]() tagged posts are off-limits to jokes or irrelevant replies. more >ĪskReddit is not your soapbox, personal army, or advertising platform. Posting, or seeking, any identifying personal information, real or fake, will result in a ban without a prior warning. more >ĪskReddit is for open-ended discussion questions. more >Īny post asking for advice should be generic and not specific to your situation alone. Your thoughts/responses to the question can go in the comments section. The title may contain two, short, necessary context sentences. You must post a clear and direct question in the title.
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