3.A.27. The Endoplasmic Reticulum Membrane Protein Insertion Complex (EMC) Family
Guna et al. 2018 found that known membrane insertion pathways fail to effectively engage tail-anchored membrane proteins with moderately hydrophobic C-terminal transmembrane domains. These proteins are instead shielded in the cytosol by calmodulin. Dynamic release from calmodulin allowed sampling of the endoplasmic reticulum (ER), where the conserved ER membrane protein complex (EMC) was shown to be essential for efficient insertion in vitro and in cells. Purified EMC in synthetic liposomes catalyzed the insertion of its substrates in a reconstituted system. Thus, EMC is a transmembrane domain insertase, a function that may explain its widely pleiotropic membrane-associated phenotypes across organisms (Guna et al. 2018). This family can be considered to be the second C-terminal Tail-Anchored Membrane Protein Biogenesis/Insertion Complex Family, the first having TC# 3.A.21. This system has been reviewed (Volkmar and Christianson 2020).
The ten known proteins of the EMC are EMC1 - EMC10. These proteins have sizes between 110 and 993 aas, seven of them having sizes between 180 and 300 aas in humans (see 3.A.27.1.1). These proteins have between ) and 3 TMSs each. None of them are clearly homologous to any of the proteins in TCDB based on sequence similarity. However, Guna et al. 2018 reported that EMC3 is distantly related to the yeast Get1 protein and the mammalian TRC40 protein, subunits of the insertase for the TRC pathway (Wang et al. 2014) (see TC#s 3.A.19 and 3.A.21). Get1 and EMC3 may have evolved from the ancestral prokaryotic insertase of the YidC family as claimed by Guna et al., 2018, The EMC has been genetically implicated in several membrane-associated processes such as quality control, trafficking, protein maturation, and lipid homeostasis (Jonikas et al. 2009; Christianson et al. 2011).
High-throughput genetic interaction analyses have revealed the EMC, and its disruption has since been found to affect wide-ranging processes, including protein trafficking, organelle communication, ER stress, viral maturation, lipid homeostasis, integral membrane insertion and folding, and others (Chitwood and Hegde 2019). Biochemical reconstitution experiments showed that EMC can directly mediate the insertion of TMSs into the lipid bilayer. A central role for EMC as a TMD insertion factor explains its high abundance, wide conservation, and pleiotropic phenotypes. Tian et al. 2019 identified 36 EMC-dependent membrane proteins and 171 EMC- independent membrane proteins, and found that a common feature among EMC-dependent proteins is the occurance of TMSs with polar and/or charged residues; the EMC may be involved in handling TMDs with residues challenging for membrane integration.
Hiramatsu et al. 2019 investigated the EMC function using the Drosophila photoreceptor as a model system. EMC was necessary only for the biogenesis of a subset of multi-pass membrane proteins such as rhodopsin (Rh1), TRP, TRPL, Csat, Cni, SERCA, and Na+K+ATPase alpha, but not for that of secretory or single-pass membrane proteins. In EMC-deficient cells, Rh1 was translated to its C-terminus but degraded independently from ER-associated degradation. Thus, EMC exerts its effect after translation but during the membrane integration of TMDs. EMC is not required for the stable expression of the first three TMSs of Rh1 but is required for that of the fourth and fifth TMSs. EMC is required for the ER membrane insertion of succeeding TMDs of multi-pass membrane proteins (Hiramatsu et al. 2019).
The reaction belived to be catalyzed by the EMC complex is:
C-terminal tail anchored protein with a moderately hydrophobic TMS (associated with calmodulin in the cytoplasm) → C-terminal tail anchored protein with a moderately hydrophobic TMS (anchored to the ER membrane).