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KMID : 0624620090420060367
BMB Reports
2009 Volume.42 No. 6 p.367 ~ p.372
Biochemical characterization of the lipid-binding properties of a broccoli cuticular wax-associated protein, WAX9D, and its application
Ahn Sun-Young

Pyee Jae-Ho
Kim Jong-Min
Park Heon-Yong
Abstract
In this study, we showed that WAX9D, a nonspecific lipid-transfer protein found in broccoli, binds palmitate (C16) and stearate (C18) with dissociation constants of 0.56 ¥ìM and 0.52 ¥ìM, respectively. WAX9D was fused to thioredoxin protein by genetic manipulation to enhance its solubility. The data revealed strong interaction of Trx-WAX9D with palmitate and stearate. The dissociation constants of Trx-WAX9D for palmitate and stearate were 1.1 ¥ìM and 6.4 ¥ìM, respectively. The calculated number of binding sites for palmitate and stearate was 2.5 to 2.7, indicating that Trx-WAX9D can bind three molecules of fatty acids. Additionally, Trx-WAX9D was shown to inhibit the apoptotic effect of palmitate in endothelial cells. Our data using Trx-WAX9D provide insight into the broad spectrum of its biological applications with specific palmitate binding.
KEYWORD
Apoptosis, Binding, Fatty acids, Fluorescence spectroscopy, Lipid-transfer proteins
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