Concanavalin A (ConA) isolated from jack bean (Canavalia ensiformis) belongs to the legume lectin family. 1)Among the monosaccharides, methyl a-D-mannopyranoside (Me a-Man, see Chart) exhibits the most potent inhibitory effect on formation of the complex of ConA with dextran, [2][3][4] and the mannotriose 3,6-di-O-(a-D-mannosyl)-a-D-mannoside was found to be the most potent oligosaccharide, being more potent than Me a-Man. 5)Shinohara et al. 6,7) examined the binding of ConA with glycosidase-treated fetuins containing the branched mannotriose moieties, which were immobilized in the chip of an optical biosensor utilizing the surface plasmon resonance (SPR) technique. [6][7][8][9] Their results showed that the terminal mannotriose moiety of glucosaminidase-treated fetuin (aglucosaminofetuin) was most sensitive. Galactosidase-treated fetuin (agalactosylfetuin) and sialidase-treated fetuin (asialofetuin, ASF) having glycosylated non-terminal mannotriose moieties bound more weakly than aglucosaminofetuin to ConA. 6)To understand the structural requirements of sugars for binding with ConA in biomembranes, we quantitatively analyzed the effects of various sugars on the dissociation of ConA from ASF using an optical biosensor system. As ASF does not interact with ConA tightly, 7) the ASF-ConA system should be favorable for determination of the affinity of sugars for ConA. In fact, we found that our system is a good mode of interaction of sugars with ConA in biomembranes. ExperimentalMaterials Human and bovine fresh blood was obtained from the Blood Center of the Japan Red Cross Society and the Meat Inspection Facility, Tokushima, respectively. ConA and ASF were purchased from Sigma. Other chemicals and reagents were of the highest grade commercially available.Preparation of the ASF-Immobilized Dextran Matrix on the Biosensor Chip SPR measurements were performed with a BIACORE 1000 (Pharmacia Biosensor AB, Uppsala, Sweden).9) We used a CM5 biosensor chip coated with carboxymethyldextran (CMD) matrix, and it was mounted in a flow-cell. The flow-cell was equilibrated with Hepes buffered saline (HBS) containing 90 mM NaCl and 20 mM Hepes/NaOH buffer (pH 7.0) at 25°C, and CMD was activated with a mixture of 200 mM N-ethyl-NЈ-(diethylaminopropyl)-carbodiimide and 50 mM N-hydroxysuccinimide. Into the activated flow-cell 30 mg/ml ASF in 10 mM sodium acetate buffer (pH 4.7) was injected. Then, free activated carboxyl groups were masked by injection of 1 M ethanolamine at pH 8.5. Each sample was injected into the ASF-immobilized flow-cell. The net amount of the conjugated ASF was calculated by subtracting its amount in the blank flow-cell from that of the ASF-immobilized flow-cell, in terms of SPR response. The immobilization of ASF to a CMD matrix in the flow-cell increased SPR by 2300 units, which may correspond to 2.3 ng/mm 2 ASF.10) The ASF-immobilized biosensor chip was stored at 4°C in the chemically stable package.Inhibition of Sugars on the Binding of Con A to Immobilized ASF A solution of 50 mg/ml ConA in HBS was injecte...

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Cannabis sativa L. is an annual herb oldest cultivated plants as a source of fiber since about 5000 B.C. On the other hand, the cannabis flower and seed are listed in Shennong’s classic Materia Medica approximately 2000 years ago. The formulas prescribed with cannabis in Kampo medicine have been summarized. Cannabidiol (CBD) and tetrahydrocannabinol (THC) are the major neurological and psychiatric cannabinoids, and develop to drugs. It becomes evident that the therapeutic CBD and/or THC are the important candidate of anti-dementia drugs having different mechanism for Alzheimer’s patients. Two receptors and endocannabinoids are also discussed for underlying mechanism of action. In order to promote the breeding of cannabis plant containing higher concentration of target cannabinoid the biosynthetic enzymes were isolated, cloning and the tertiary structure of THCA synthase determined by x-ray analysis resulting in the possibility of molecular breeding for cannabinoids.

2014J o u r n a l o f I n f o r m a t i o n P r o c e s s i n g a n d M a n a g e m e n t JOHO KANRIJuly