title: Expression and characterization of human lamin C.
abstract: We have expressed human lamin C cDNA in E. coli using a modification of the pLcII vector system. Protein produced in this way had seven additional amino acids at its N-terminus, but retained key lamin structural and assembly properties. The modified vector we produced may prove useful when difficulties are encountered in removal of the cII fusion peptide by factor X cleavage in the pLcII system. Shadowed preparations of expressed lamin C showed the presence of 50-nm rod-like particles that closely resembled those observed for native material. Isolated molecules had two globular domains at one end, indicating that they were constructed from two parallel polypeptide chains. The expressed material also formed paracrystals with a characteristic 22.5 nm axial repeat, indicating that its assembly properties had also been retained. We also used site-specific mutagenesis to engineer a lamin fragment that lacked the C-terminal non-helical domain of the molecule. This material formed paracrystals similar to those obtained with the intact molecule, indicating that the large C-terminal non-helical domain did not contain information vital for lamin assembly.
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge, UK.
authors.forename: R D
authors.initials: RD
authors.lastname: Moir
authors.forename: R A
authors.initials: RA
authors.lastname: Quinlan
authors.forename: M
authors.initials: M
authors.lastname: Stewart
mesh_terms: D003001:Cloning, Molecular; D003461:Crystallography; D015870:Gene Expression; D006801:Humans; D034904:Lamin Type A; D034882:Lamins; D008854:Microscopy, Electron; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000032:analysis* / Q000235:genetics / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins; D011994:Recombinant Proteins; C440308:lamin C