Expression, purification and microscopic characterization of human ATP-binding cassette sub-family B member 7 protein

• Published in: Protein expression and purification Vol. 183; p. 105860
• Main Authors: Nie, Zhenzhen, Yan, Qinqin, Shen, Yuequan, Yang, Xue
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2021
• Subjects: 2D classification; ABCB7; Anemia, Sideroblastic - enzymology; Anemia, Sideroblastic - genetics; ATP-Binding Cassette Transporters - biosynthesis; ATP-Binding Cassette Transporters - chemistry; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - isolation & purification; Electron microscopy; Expression; Genetic Diseases, X-Linked - enzymology; Genetic Diseases, X-Linked - genetics; Humans; Mutation; Protein Conformation; Purification; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Spinocerebellar Ataxias - enzymology; Spinocerebellar Ataxias - genetics; Purification; Electron microscopy; ABCB7; Expression; 2D classification
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2021.105860

The ATP-binding cassette sub-family B member 7 (ABCB7) is a membrane transport protein located on the inner membrane of mitochondria, which could be involved in the transport of heme from the mitochondria to the cytosol. ABCB7 also plays a central role in the maturation of cytosolic iron-sulfur (Fe/S) cluster-containing proteins, and mutations can cause a series of mitochondrial defects. X-linked sideroblastic anemia and ataxia (XLSA-A) is a rare cause of early onset ataxia, which may be overlooked due to the usually mild asymptomatic anemia. The genetic defect has been identified as a mutation in the ABCB7 gene at Xq12-q13. Here, we report the expression, purification and the 2D projections derived from negatively stained electron micrographs of recombinant H. sapiens ABCB7 (hABCB7), paving the way from an atomic structure determination of ABCB7. •The full-length of human ABCB7 transporter was expressed using pEG BacMam. system.•The human ABCB7 protein was purified with affinity and size-exclusion chromatography.•Electron microscopic characterization lays the foundation for future structural and. functional investigation of ABCB7.