title: Identification and characterization of SlVKOR, a disulfide bond formation protein from Solanum lycopersicum, and bioinformatic analysis of plant VKORs.
abstract: Homologs of vitamin K epoxide reductase (VKOR) exist widely in plants. However, only VKOR of Arabidopsis thaliana has been the subject of many studies to date. In the present study, the coding region of a VKOR from Solanum lycopersicum (JF951971 in GenBank) was cloned; it contained a membrane domain (VKOR domain) and an additional soluble thioredoxin-like (Trx-like) domain. Bioinformatic analysis showed that the first 47 amino acids in the N-terminus should act as a transit peptide targeting the protein to the chloroplast. Western blot demonstrated that the protein is localized in thylakoid membrane with the Trx-like domain facing the lumen. Modeling of three-dimensional structure showed that SlVKOR has a similar conformation with Arabidopsis and cyanobacterial VKORs, with five transmembrane segments in the VKOR domain and a typical Trx-like domain in the lumen. Functional assay showed that the full-length of SlVKOR with Trx-like domain without the transit peptide could catalyze the formation of disulfide bonds. Similar transit peptides at the N-terminus commonly exist in plant VKORs, most of them targeting to chloroplast according to prediction. Comparison of sequences and structures from different plants indicated that all plant VKORs possess two domains, a transmembrane VKOR domain and a soluble Trx-like domain, each having four conservative cysteines. The cysteines were predicted to be related to the function of catalyzing the formation of disulfide bonds.
authors: College of Life Sciences, Shandong Agricultural University, Taian, Shandong, 271018, P. R. China. xyunwang@sdau.edu.cn.
authors: Chun-Mei
authors: CM
authors: Wan
authors: Xiao-Jian
authors: XJ
authors: Yang
authors: Jia-Jia
authors: JJ
authors: Du
authors: Ying
authors: Y
authors: Lu
authors: Zhi-Bo
authors: ZB
authors: Yu
authors: Yue-Guang
authors: YG
authors: Feng
authors: Xiao-Yun
authors: XY
authors: Wang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D018551:Solanum lycopersicum / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D064417:Vitamin K Epoxide Reductases / Q000737:chemistry / Q000145:classification / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D064417:Vitamin K Epoxide Reductases