GTPase activity and biochemical characterization of a recombinant cotton fiber annexin
- Univ. of Texas, Austin, TX (United States). Dept. of Botany
A cDNA encoding annexin was isolated from a cotton (Gossypium hirsutum) fiber cDNA library. The cDNA was expressed in Escherichia coli, and the resultant recombinant protein was purified. The authors then investigated some biochemical properties of the recombinant annexin based on the current understanding of plant annexins. An add-back experiment was performed to study the effect of the recombinant annexin on [beta]-glucan synthase activity, but no effect was found. However, it was found that the recombinant annexin could display ATPase/GTPase activities. The recombinant annexin showed much higher GTPase than ATPase activity. Mg[sup 2+] was essential for these activities, whereas a high concentration of Ca[sup 2+] was inhibitory. A photolabeling assay showed that this annexin could bind GTP more specifically than ATP. The GTP-binding site on the annexin was mapped into the carboxyl-terminal fourth repeat of annexin from the photolabeling experiment using domain-deletion mutants of this annexin. Northern-blot analysis showed that the annexin gene was highly expressed in the elongation stages of cotton fiber differentiation, suggesting a role of this annexin in cell elongation.
- Sponsoring Organization:
- USDOE; USDOE, Washington, DC (United States)
- DOE Contract Number:
- FG03-94ER20145
- OSTI ID:
- 6281362
- Journal Information:
- Plant Physiology; (Bethesda), Vol. 119:3; ISSN 0032-0889
- Country of Publication:
- United States
- Language:
- English
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