title: Characterization of a novel annexin gene from cotton (Gossypium hirsutum cv CRI 35) and antioxidative role of its recombinant protein.
abstract: Plant annexins represent a multigene family involved in cellular elongation and development. A cDNA encoding a novel annexin was isolated from a cotton (Gossypium hirsutum) fiber cDNA library and designated GhAnx1. This gene encodes a 316 amino acid protein with a theoretical molecular mass of 36.06 kDa and a theoretical pI of 6.19. At the amino acid level, it shares high sequence similarity and has evolutionary relationships with annexins from higher plants. The purified recombinant protein expressed in Escherichia coli was used to investigate its physicochemical properties. Circular dichroism spectrum analyses showed a positive peak rising to the maximum at 196 nm and a broad negative band rounding 215 nm, suggesting that the GhAnx1 protein was prominently α-helical. The fluorescence measurements indicated that it could bind to Ca(2+) in vitro. These results demonstrated that GhAnx1 was a typical annexin protein in cotton. A bioassay experiment was conducted to analyze its potential function and showed that E. coli cells expressing GhAnx1 were protected from tert-butyl hydroperoxide (tBH) stress, suggesting that it had a potential antioxidative role. Northern blot analyses revealed that GhAnx1 was highly expressed in fibers, especially during the elongation stage, suggesting that it might be important for fiber elongation.
authors.affiliation: Laboratory of Molecular Biology and MOE Laboratory of Protein Science, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhou
authors.forename: Jin
authors.initials: J
authors.lastname: Duan
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Wang
authors.forename: Heng-Mu
authors.initials: HM
authors.lastname: Zhang
authors.forename: Ming-Xing
authors.initials: MX
authors.lastname: Duan
authors.forename: Jin-Yuan
authors.initials: JY
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D017302:Annexins / Q000235:genetics* / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D015152:Blotting, Northern; D015870:Gene Expression; D003368:Gossypium / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D017302:Annexins; D000975:Antioxidants; D010940:Plant Proteins; D011994:Recombinant Proteins