biostudies-literatureHollinger JAmerican Heart AssociationNHLBI NIH HHSFoundation for the National Institutes of HealthNational Institutes of HealthNIGMS NIH HHS106322https://www.ebi.ac.uk/biostudies/studies/S-EPMC11143474biostudies-literatureUnknown210The protein Family with sequence similarity 210 member A (FAM210A) is a mitochondrial inner membrane protein that regulates the protein synthesis of mitochondrial DNA encoded genes. However, how it functions in this process is not well understood. Developing and optimizing a protein purification strategy will facilitate biochemical and structural studies of FAM210A. Here, we developed a method to purify human FAM210A with deleted mitochondrial targeting signal sequence using the MBP-His₁₀ fusion in Escherichia coli. The recombinant FAM210A protein was inserted into the E. coli cell membrane and purified from isolated bacterial cell membranes, followed by a two-step process using Ni-NTA resin-based immobilized-metal affinity chromatography (IMAC) and ion exchange purification. A Protein expression and purificationExpression and purification of the mitochondrial transmembrane protein FAM210A in Escherichia coli.PMC11143474R35 GM133462R01HL164584R01 HL136432R01HL169432R35GM13346R01HL132899R01 HL132899R01 HL164584R01 HL147954R01 HL16943219POST34400013R01HL147954848985Wu JHollinger JYao PAwayda KMO'Connell MRfalseExpression and purification of the mitochondrial transmembrane protein FAM210A in Escherichia coli.The protein Family with sequence similarity 210 member A (FAM210A) is a mitochondrial inner membrane protein that regulates the protein synthesis of mitochondrial DNA encoded genes. However, how it functions in this process is not well understood. Developing and optimizing a protein purification strategy will facilitate biochemical and structural studies of FAM210A. Here, we developed a method to purify human FAM210A with deleted mitochondrial targeting signal sequence using the MBP-His₁₀ fusion in Escherichia coli. The recombinant FAM210A protein was inserted into the E. coli cell membrane and purified from isolated bacterial cell membranes, followed by a two-step process using Ni-NTA resin-based immobilized-metal affinity chromatography (IMAC) and ion exchange purification. A 2023-01-01T00:00:00Z2023 Oct2026-06-02T06:54:31.969Z2026-04-15T03:15:34.224ZS-EPMC111434743732993410.1016/j.pep.2023.106322