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Insight into vitamin B 6 ‐dependent epilepsy due to PLPBP (previously PROSC ) missense mutations
Author(s) -
Tremiño Lorena,
ForcadaNadal Alicia,
Rubio Vicente
Publication year - 2018
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.23540
Subject(s) - missense mutation , biology , epilepsy , genetics , mutation , neuroscience , gene
Abstract Vitamin B 6 ‐dependent genetic epilepsy was recently associated to mutations in PLPBP (previously PROSC ), the human version of the widespread COG0325 gene that encodes TIM‐barrel‐like pyridoxal phosphate (PLP)‐containing proteins of unclear function. We produced recombinantly, purified and characterized human PROSC (called now PLPHP) and its six missense mutants reported in epileptic patients. Normal PLPHP is largely a monomer with PLP bound through a Schiff‐base linkage. The PLP‐targeting antibiotic d ‐cycloserine decreased the PLP‐bound peak as expected for pseudo‐first‐order reaction. The p.Leu175Pro mutation grossly misfolded PLPHP. Mutations p.Arg241Gln and p.Pro87Leu decreased protein solubility and yield of pure PLPHP, but their pure forms were well folded, similarly to pure p.Pro40Leu, p.Tyr69Cys, and p.Arg205Gln mutants (judged from CD spectra). PLPHP stability was decreased in p.Arg241Gln, p.Pro40Leu, and p.Arg205Gln mutants (thermofluor assays). The p.Arg241Gln and p.Tyr69Cys mutants respectively lacked PLP or had a decreased amount of this cofactor. With p.Tyr69Cys there was extensive protein dimerization due to disulfide bridge formation, and PLP accessibility was decreased (judged from d ‐cycloserine reaction). A 3‐D model of human PLPHP allowed rationalizing the effects of most mutations. Overall, the six missense mutations caused ill effects and five of them impaired folding or decreased stability, suggesting the potential of pharmacochaperone‐based therapeutic approaches.