title: Expression, purification and crystallization of the SKICH domain of human TAX1BP1.
abstract: TAX1BP1 is a highly conserved, pleiotropic protein that plays many essential functions in human cells, including negative regulation of inflammatory and antimicrobial responses mediated by NF-κB and IRF3 signaling, inhibition of apoptosis, transcriptional coactivation and autophagy etc. TAX1BP1 contains a SKICH domain at the N-terminus, three coiled-coil domains in the middle and two ubiquitin-binding zinc-finger motifs at the C-terminus. The SKICH domain and the linker sequence between the SKICH domain and the coiled-coil region mediate interaction with ubiquitin-like proteins of the LC3/GABARAP family, which are autophagosome markers. For structure determination of the SKICH domain of TAX1BP1, a protein construct (amino acids 15-148) corresponding to the SKICH domain plus the linker region was expressed, purified and crystallized. A native diffraction data set has been collected to 1.9 Å resolution. A molecular-replacement solution has been found by using the structure of the SKICH domain of NDP52, a paralog of TAX1BP1.
authors.affiliation: Department of Chemistry and Biochemistry, Southern Illinois University, 1245 Lincoln Drive, Carbondale, IL 62901, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Chemistry and Biochemistry, Southern Illinois University, 1245 Lincoln Drive, Carbondale, IL 62901, USA.
authors.forename: Guan
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Computer Science, Southern Illinois University, 1245 Lincoln Drive, Carbondale, IL 62901, USA.
authors.forename: Xiaolan
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Chemistry and Biochemistry, Southern Illinois University, 1245 Lincoln Drive, Carbondale, IL 62901, USA.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Du
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D003460:Crystallization; D005786:Gene Expression Regulation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis* / Q000302:isolation & purification; D017434:Protein Structure, Tertiary / Q000502:physiology
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; C120635:TAX1BP1 protein, human; C471548:SKIP enzyme, human; D010744:Phosphoric Monoester Hydrolases
keywords: A20; IRF3; NF-κB; SKICH domain; TAX1BP1; autophagy receptor; innate immune signaling