2009年7月
Purification and biochemical characterization of a D-galactose binding lectin from Japanese sea hare (Aplysia kurodai) eggs
BIOCHEMISTRY-MOSCOW
- 巻
- 74
- 号
- 7
- 開始ページ
- 709
- 終了ページ
- 716
- 記述言語
- 英語
- 掲載種別
- DOI
- 10.1134/S0006297909070025
- 出版者・発行元
- MAIK NAUKA/INTERPERIODICA/SPRINGER
A lectin was purified from Japanese sea hare Aplysia kurodai by lactosyl-agarose affinity chromatography. The molecular mass of the lectin was determined to be 56 and 32 kDa by SDS-PAGE under non-reducing and reducing conditions, respectively. It was found to agglutinate trypsinized and glutaraldehyde-fixed rabbit and human erythrocytes in the absence of divalent cations. The lectin exhibited stable thermo-tolerance as it retained hemagglutinating activity for 1 h even at 80A degrees C and showed stability at pH 10. By contrast, it was very sensitive at pH less than 5 and in the presence of the sulfhydryl-group preserving reagent, beta-mercaptoethanol. The hemagglutinating activity by the lectin was specifically inhibited by D-galactose, galacturonic acid, methyl-alpha- and methyl-beta-D-galactopyranoside, lactose, melibiose, and asialofetuin. The association rate constant (k (ass)) and dissociation rate constant (k (diss)) were determined for the lectin to be 4.3 center dot 10(5) M-1 center dot sec(-1) and 2.2 center dot 10(-3) sec(-1), respectively, using a surface plasmon resonance biosensor. The lectin moderately inhibited cell proliferation in the P388 cell line dose dependently. Interestingly, lectin-treated cells did not show a fragmented DNA ladder as is caused by apoptosis, suggesting that the cell proliferation inhibition was caused by another unknown mechanism.
- リンク情報
- ID情報
-
- DOI : 10.1134/S0006297909070025
- ISSN : 0006-2979
- eISSN : 0320-9725
- Web of Science ID : WOS:000268256500002