New insights into the pathogenicity of TMEM165 variants using structural modeling based on AlphaFold 2 prediction

MEM165 is a Golgi protein playing a crucial role in Mn²⁺ transport, and whose mutations in patients are known to cause Congenital Disorders of Glycosylation. Some of those mutations affect the highly-conserved consensus motifs E-φ-G-D-[KR]-[TS] characterizing the CaCA2/UPF0016 family, presumably important for the transport of Mn²⁺ which is essential for the function of many Golgi glycosylation enzymes. Others, like the G>R³⁰⁴ mutation, are far away from these motifs in the sequence. Until recently, the classical membrane protein topology prediction methods were unable to provide a clear picture of the organization of TMEM165 inside the cell membrane, or to explain in a convincing manner the impact of patient and experimentally-generated mutations on the transporter function of TMEM165. In this study, AlphaFold 2 was used to build a TMEM165 model that was then refined by molecular dynamics simulation with membrane lipids and water.