5IO9

X-RAY STRUCTURE OF THE N-TERMINAL DOMAIN OF HUMAN DOUBLECORTIN

5IO9 の概要

• エントリーDOI: 10.2210/pdb5io9/pdb
• 分子名称: Neuronal migration protein doublecortin (2 entities in total)
• 機能のキーワード: dcx domain, ubiquitin-like fold, microtubule associated, signaling protein, transferase
• 由来する生物種: Homo sapiens (Human)
• 細胞内の位置: Cytoplasm : O43602
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 24409.08

構造登録者

Ruf, A.,Benz, J.,Burger, D.,D'Arcy, B.,Debulpaep, M.,Di Lello, P.,Fry, D.,Huber, W.,Kremer, T.,Laeremans, T.,Matile, H.,Ross, A.,Rudolph, M.G.,Rufer, A.C.,Sharma, A.,Steinmetz, M.O.,Steyaert, J.,Schoch, G.,Stihle, M.,Thoma, R. (登録日: 2016-03-08, 公開日: 2016-03-23, 最終更新日: 2024-01-10)

主引用文献

Burger, D.,Stihle, M.,Sharma, A.,Di Lello, P.,Benz, J.,D'Arcy, B.,Debulpaep, M.,Fry, D.,Huber, W.,Kremer, T.,Laeremans, T.,Matile, H.,Ross, A.,Rufer, A.C.,Schoch, G.,Steinmetz, M.O.,Steyaert, J.,Rudolph, M.G.,Thoma, R.,Ruf, A.
Crystal Structures of the Human Doublecortin C- and N-terminal Domains in Complex with Specific Antibodies.
J.Biol.Chem., 291:16292-16306, 2016

PubMed Abstract: Doublecortin is a microtubule-associated protein produced during neurogenesis. The protein stabilizes microtubules and stimulates their polymerization, which allows migration of immature neurons to their designated location in the brain. Mutations in the gene that impair doublecortin function and cause severe brain formation disorders are located on a tandem repeat of two doublecortin domains. The molecular mechanism of action of doublecortin is only incompletely understood. Anti-doublecortin antibodies, such as the rabbit polyclonal Abcam 18732, are widely used as neurogenesis markers. Here, we report the generation and characterization of antibodies that bind to single doublecortin domains. The antibodies were used as tools to obtain structures of both domains. Four independent crystal structures of the N-terminal domain reveal several distinct open and closed conformations of the peptide linking N- and C-terminal domains, which can be related to doublecortin function. An NMR assignment and a crystal structure in complex with a camelid antibody fragment show that the doublecortin C-terminal domain adopts the same well defined ubiquitin-like fold as the N-terminal domain, despite its reported aggregation and molten globule-like properties. The antibodies' unique domain specificity also renders them ideal research tools to better understand the role of individual domains in doublecortin function. A single chain camelid antibody fragment specific for the C-terminal doublecortin domain affected microtubule binding, whereas a monoclonal mouse antibody specific for the N-terminal domain did not. Together with steric considerations, this suggests that the microtubule-interacting doublecortin domain observed in cryo-electron micrographs is the C-terminal domain rather than the N-terminal one.

PubMed: 27226599
DOI: 10.1074/jbc.M116.726547

実験手法

X-RAY DIFFRACTION (1.3 Å)