Genes contain the instructions needed to make proteins from smaller building blocks called amino acids. These instructions are first transcribed to produce molecules of messenger RNA, which are then translated by a ribosome. This ‘molecular machine’ translates the instructions in the messenger RNA into the sequence of amino acids needed to make the protein. For some proteins to carry out their role, they need to be delivered to the outside of the cell, or inserted into one of the cell's membranes. As they are being built, these proteins are identified by a so-called ‘signal recognition particle’, which is often called an SRP for short. The SRP attaches to the new protein when it is still joined to the ribosome, and pulls the protein-ribosome complex to an opening in the target membrane. The new protein chain then enters this opening and either passes through to the other side of the membrane, or ends up embedded within it. To date, most studies that have investigated this process have involved scientists stalling the building of the new protein to see how SRP interacts with inactivated protein-ribosome complexes. Unfortunately, this means that some of the details of what happens during this process have likely been missed. Now, Noriega et al. have addressed this problem by developing a method to watch, in real-time, a single active protein-ribosome complex interacting with individual SRPs. This was achieved by attaching fluorescent molecules to SRP and protein-ribosome complexes purified from the bacterium E. coli. The distance between the two fluorescent molecules was then tracked over time. This revealed that the SRP typically binds to the protein-ribosome complex after 40–55 amino acids have been built into the protein. At this point, a so-called ‘signal sequence’ of amino acids has emerged from the complex and can be recognized by the SRP. Earlier studies had suggested that signal sequences might tell the SRP when to bind, but this had not been demonstrated in experiments using active protein-ribosome complexes. The strategy of using fluorescent molecules to follow single molecules undergoing this process in real-time could now be used by other scientists to re-examine and determine new properties of the protein-ribosome complex in action.
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