There are two E1, ∼50 E2, and ∼500 E3 enzymes in the human genome; thus the substrate specificity of ubiquitination is mainly determined by different combinations of E2–E3 complexes (Ciechanover, 2006). E3 enzymes can add a single ubiquitin molecule to the acceptor lysine residue of the substrate (monoubiquitination) or they can add ubiquitin monomers sequentially to form a polyubiquitin chain (Nagy and Dikic, 2010). Monoubiquitination does not signal for

proteasomal degradation but rather seems to regulate protein trafficking and other processes. The outcome of polyubiquitination depends on which lysine residue of the seven present in ubiquitin selleck chemical is utilized for constructing the chain. Lysine-48 (K48)-linked polyubiquitin chains target

proteins for proteasomal degradation, whereas K63 chains are Everolimus purchase used for nonproteasomal functions such as protein kinase activation, regulation of protein-protein interactions, and control of receptor endocytosis (Nagy and Dikic, 2010). By utilizing different lysine residues, the ubiquitination system can generate diverse polyubiquitin structures and varied signaling outcomes, which are still not fully understood in neurons or other cell types. Once a substrate is ubiquitinated by K48 chains, it is conveyed to the 26S proteasome by E3s themselves, substrate-shuttling factors, or binding to resident polyubiquitin receptors on the proteasome (Glickman and Raveh, 2005). Both in neurons and nonneuronal cells, proteasome activity and subcellular localization mafosfamide can be dynamically modulated through posttranslational modifications and regulated interactions with accessory proteins, such as CaMKIIα (Bingol and Schuman, 2006, Bingol et al., 2010, Djakovic et al., 2009 and Glickman and Raveh, 2005). There is also evidence for different proteasome-interacting proteins in brain versus other tissues and even between synaptic versus cytosolic compartments within neurons, suggesting proteasome heterogeneity across cell types and subcellular compartments (Tai et al., 2010). Protein ubiquitination is a dynamic and reversible process owing to the action of deubiquitinating enzymes (DUBs; ∼100

in the human genome) (Komander et al., 2009). DUBs can both facilitate and antagonize ubiquitin-mediated signaling and protein degradation. They promote ubiquitination in general by providing free ubiquitin through cleavage of ubiquitin monomers from polyubiquitin chains. On the other hand, DUBs counteract the function of E3 ligases and stabilize proteins by removing ubiquitin from substrates before they can be destroyed by the proteasome. DUBs can also remove monoubiquitin and other types of polyubiquitin linkages (such as K63-polyubiquitin) to terminate proteasome-independent ubiquitin signaling (Komander et al., 2009). To date, several ubiquitin conjugation and removal enzymes have been described that regulate synaptic function (see Table 1 and Table 2 and Figure 1).