title: Mth10b, a unique member of the Sac10b family, does not bind nucleic acid.
abstract: The Sac10b protein family is regarded as a group of nucleic acid-binding proteins that are highly conserved and widely distributed within archaea. All reported members of this family are basic proteins that exist as homodimers in solution and bind to DNA and/or RNA without apparent sequence specificity in vitro. Here, we reported a unique member of the family, Mth10b from Methanobacterium thermoautotrophicum ΔH, whose amino acid sequence shares high homology with other Sac10b family proteins. However, unlike those proteins, Mth10b is an acidic protein; its potential isoelectric point is only 4.56, which is inconsistent with the characteristics of a nucleic acid-binding protein. In this study, Mth10b was expressed in Escherichia coli and purified using a three-column chromatography purification procedure. Biochemical characterization indicated that Mth10b should be similar to typical Sac10b family proteins with respect to its secondary and tertiary structure and in its preferred oligomeric forms. However, an electrophoretic mobility shift analysis (EMSA) showed that neither DNA nor RNA bound to Mth10b in vitro, indicating that either Mth10b likely has a physiological function that is distinct from those of other Sac10b family members or nucleic acid-binding ability may not be a fundamental factor to the actual function of the Sac10b family.
authors.affiliation: Ministry of Education Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, People's Republic of China.
authors.forename: Yan-Feng
authors.initials: YF
authors.lastname: Liu
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Yao
authors.forename: Xian-Ming
authors.initials: XM
authors.lastname: Pan
authors.forename: Meng
authors.initials: M
authors.lastname: Ge
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000378:metabolism*; D002853:Chromatography, Liquid; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D019843:Archaeal Proteins; D009696:Nucleic Acids