Determining composition of micron-scale protein deposits in neurodegenerative disease by spatially targeted optical microproteomics

Neurodegenerative diseases such as Alzheimer's disease affect millions of people worldwide. In many of these diseases, toxic proteins accumulate in the brain and build up as small ‘plaques’ in the gaps, or synapses, that cells called neurons communicate across. Eventually, the plaques prevent the neurons signaling to each other correctly, leading to problems such as memory loss. Identifying the proteins present in plaques is technically challenging, partly because the plaques are very small. Hadley, Rakhit et al. have now developed a new method called spatially targeted optical microproteomics (or STOMP) that can collect proteins from small areas of cells. In this method, plaques are identified under a light microscope, and their contents are attached to a molecule called a photo-affinity tag using lasers. The photo-tagged proteins are then pulled out using beads that specifically bind to the photo-affinity tag. The proteins can then be identified using a well-established method called mass spectrometry. Hadley, Rakhit et al. used STOMP to analyze plaques present in the brains of mice that develop similar symptoms to those seen in humans with Alzheimer's disease. This revealed that these plaques contain more than 50 different proteins, some of which had not previously been found in plaques. In particular, several proteins from the ‘presynaptic’ neuron that sends signals across the synapse were found in the plaques. However, no proteins from the receiving (‘postsynaptic’) neuron on the other side of the synapse were present in the plaque. Fixed human brain tissue is more difficult to analyze than mouse samples because it is modified for storage. In spite of these issues, Hadley, Rakhit et al. successfully also used STOMP to identify the proteins in human plaques. STOMP can be used to identify the proteins present in any area of a cell and thus has the potential to be widely used by scientists, not just those studying plaques.