Structural studies of KCTD1 and its disease-causing mutant P20S provide insights into the protein function and misfunction

• Published in: International journal of biological macromolecules 2024-10, Vol.277 (Pt 4), p.134390, Article 134390
• Main Authors: Balasco, Nicole, Ruggiero, Alessia, Smaldone, Giovanni, Pecoraro, Giovanni, Coppola, Luigi, Pirone, Luciano, Pedone, Emilia M., Esposito, Luciana, Berisio, Rita, Vitagliano, Luigi
• Format: Article
• Language: English
• Subjects: Domain swapping; KCTD proteins; Scalp-ear-nipple syndrome
• ISSN: 0141-8130; 1879-0003; 1879-0003
• DOI: 10.1016/j.ijbiomac.2024.134390

Members of the KCTD protein family play key roles in fundamental physio-pathological processes including cancer, neurodevelopmental/neuropsychiatric, and genetic diseases. Here, we report the crystal structure of the KCTD1 P20S mutant, which causes the scalp-ear-nipple syndrome, and molecular dynamics (MD) data on the wild-type protein. Surprisingly, the structure unravels that the N-terminal region, which precedes the BTB domain (preBTB) and bears the disease-associated mutation, adopts a folded polyproline II (PPII) state. The KCTD1 pentamer is characterized by an intricate architecture in which the different subunits mutually exchange domains to generate a closed domain swapping motif. Indeed, the BTB of each chain makes peculiar contacts with the preBTB and the C-terminal domain (CTD) of an adjacent chain. The BTB-preBTB interaction consists of a PPII-PPII recognition motif whereas the BTB-CTD contacts are mediated by an unusual (+/−) helix discontinuous association. The inspection of the protein structure, along with the data emerged from the MD simulations, provides an explanation of the pathogenicity of the P20S mutation and unravels the role of the BTB-preBTB interaction in the insurgence of the disease. Finally, the presence of potassium bound to the central cavity of the CTD pentameric assembly provides insights into the role of KCTD1 in metal homeostasis.