Biological Characteristics of Serum Agglutinin from Portunus trituberculatus

The agglutination characteristics of serum agglutinin from Portunus trituberculatus were studied in this paper. The results showed that the serum agglutinin had no agglutination to crucian carp, Chinese soft shell turtle, grass carp, chicken or human group A, B or O blood cells, but had strong agglutination to blood cells of mice and rabbits. The activities of serum agglutinin from Portunus trituberculatus to mice blood cells reached 210, and to rabbit blood cells reached 28. Salinity had a greater effect on agglutinin activity. When the Na Cl concentration exceeded 0.6 mol/L,the serum agglutinin from Portunus trituberculatus was basically inactivated. The optimum p H for agglutinin activity was 6.0-7.4. The serum agglutinin from Portunus trituberculatus had obvious dependence to Ca2 ＋and Mg2 ＋, and EDTA could significantly inhibit its activity. The results of sugar inhibition test showed that the activity of agglutinin from Portunus trituberculatus can be specifically inhibited by N-acetylglucosamine and N-acetylmannosamine. The serum agglutinin from Portunus trituberculatus was isolated by ammonium sulfate gradient precipitation, and its activity was highest by the 25% ammonium sulfate precipitation system. The SDS polyacrylamide gel electrophoresis（SDS-PAGE） showed that the protein bands were mainly distributed within 72-95 ku.

Purification, Biological Activities, and Molecular Cloning of a Novel Mannose-Binding Lectin from Bulbs of Zephyranthes candida Herb （Amaryllidaceae）

A novel mannose-bindlng aggiutinln was purified from bulbs of Zephyranthes candida Herb by extraction, precipitation with 80% （NH4）2SO4, and ion-exchange chromatography on DEAE-Sepharose followed by gel flitration on Sephscryl S-100. The purified Z. candida agglutlnln （ZCA） migrated as a single band of 12 kDa on sodium dodecyi suifate-poiyecryiamide gel electrophoresis under reducing and non-reducing conditions. The apparent molecular mass of the iectln, as datermlned by gel filtration chromatography, was 48 kDa. The results Indicated that ZCA was composed of four Identical subunlts of 12 kDa each （homotetramerlc nature）. The ZCA agglutlhated rabbit erythrocytes, Escherichla coil and Saccharomyces cerevislae ceils at concentrations of 0.95, 1.90, and 31.30 μg/mL, respectively. Bloassays Indicated that ZCA has a significant effect on wheat aphid survival. Mortality after 7 d was 〉 90% at 0.26%. A degenerate primer was designed In accordance with the N-terminal partial sequence of purified ZCA. The full-length cDNA was cloned by 3＇- and 5＇-rapid amplification of cDNA ends. The full-length cDNA had 661 bp and the sequence encoded an open reading frame of 168 amino acids. The mature protein of ZCA Includes 109 amino acid residues and the molecular weight of the protein was 12.1 kDa. The result show that the zca gene encodes a protein precursor with a signal peptlde, a mature protein, and a C-terminal cleavage amino acids sequence. Molecular modeling of ZCA Indicated that Its three-dimensional atructure strongly resembies that of the snowdrop aggiutinin. Blocks＇ analysis revealed that the deduced amino acid sequence of ZCA has three functional domains specific for agglutination and three carbohydrate binding boxes （QDNY）.