Purification and properties of thiamine-binding proteins from sesame seed

Three thiamine-binding proteins of 17-19 kDa (STBP-I, II, and III) were purified from sesame seed (Sesamum indicum L.). Each of the proteins was composed of two subunits of equal molecular mass and each subunit consisted of a large polypeptide and a small polypeptide linked by a disulfide bond(s). They were rich in glutamic acid (or glutamine) and arginine. Their binding activities were optimal at neutral pH. They bound specifically free thiamine but not thiamine phosphates. STBP-I had higher affinity for thiamine than STBP-II or STBP-III. STBP-II and STBP-III bound one molecule of thiamine per molecule, and STBP-I bound 0.5 molecule. The amino acid composition and structure of the STPBs were similar to those of 2S storage proteins.     

A sulfotransferase model: Covalent participation of a macrocyclic oxime in the hydrolysis of 2,4-dinitrophenyl sulfate

The liberation of 2,4-dinitrophenolate ion from 2,4-dinitrophenyl sulfate (DNPS) in aqueous organic solvent with 0.1 N sodium hydroxide was accelerated upon addition of an equimolar amount of Oxime-I (10-hydroxy-11-hydroxyimino[20]-paracyclophane) to the sulfate ester. Oxime-I was found to undergo covalent participation at the oxime group to afford oxime O-sulfonate. The rate acceleration with Oxime-I was larger than that with β-CD (cycloheptaamylose). The catalytic efficiency of Oxime-I has been ascribed primarily to the tighter inclusion of the substrate ester into the more hydrophobic Oxime-I cavity provided by the effective apolar paracyclophane skeleton, as well as to the greater nucleophilicity of the oxime group than of the hydroxyl group in β-CD. Consequently, Oxime-I may be considered as a conventional model for arylsulfatases and sulfotransferases, providing the effective binding process for the substrate.     

A quantitative approach to understand the ultramicro-structure responsible for input mechanism in excitation-contraction (E-C) coupling, on the basis of physio-morphological observations]

Our morphophysiological studies using concanavalin A-ferritin (Con A-F) have indicated that: (1) an out- and up-ward movement of a movable structure at the luminal surface-portion of the T-tubular membrane opposite the feet initiates contraction; (2) the grade of the movement depends on that of depolarization; (3) the movable structure is essentially a 'moving arm', which is fixed in wall of T-tubules at its fixed end and is able to be bound to the Con A-moiety of Con A-F particle about at its free end. Calculation based on molecular morphology and behaviour of Con A-F particle revealed following points: If (a) the origin of coordinate be the intersection of longitudinal center line of foot and the surface of T-tubular membrane in the transverse section of the tubules, (b) the fixed point of the arm is exactly on the surface of T-tubular membrane, and (c) the movement takes place in the transverse direction to the longitudinal axis of T-tubules, (1) the location of the center point of the movement of the moving arm is at 5.4 nm in the outside direction from the origin, (2) the arm is about 4 nm in length and moves by about 2.4 nm up- and out-ward at its free end upon about complete depolarization.     

Presence of histamine H2-receptors on human gastric carcinoma cell line MKN-45 and their increase by retinoic acid treatment.

Histamine dose-dependently stimulated cyclic AMP production in human gastric carcinoma cell line MKN-45, and this effect was inhibited by cimetidine but not by pyrilamine. Moreover, not only histamine but also cimetidine displaced the specific binding of [3H]tiotidine to these cells, whereas pyrilamine had no effect. On the other hand, pretreatment of MKN-45 cells with retinoic acid (RA) significantly enhanced histamine-induced increase of cyclic AMP production, although the cyclic AMP response to either forskolin or NaF was not affected. Finally, RA treatment increased the number of histamine receptor without altering its affinity. Thus, it appears that histamine H2-receptors are present on MKN-45 cells, and that RA treatment enhances the action of histamine on these cells by increasing the number of H2-receptors.     

Observations on the effect of visual and olfactory ablation on the swimming behavior of migrating adult chum salmon,Oncorhynchus keta

The behavior of chum slamon,Oncorhynchus keta, was studied using ultrasonic telemetry in the waters off the Okhotsk coast of Hokkaido from 1979 to 1981. Thirty-six adult fish were outfitted with a 50 KHz ultrasonic transmitter provided with either a depth sensor, depth/illumination sensors, or depth/ temperature sensors. Twenty-one of the experimental fish were used as controls and left intact. Of the remaining fish, six had their sight destroyed, seven had their olfactory nerves severed or their nares filled with wax, one had both senses destroyed, and one had a sham operation, but had only a shallow transverse cut made in the skin over the olfactory nerves. Control fish and the fish with the sham operation initially swam in a horizontal zigzag pattern while fish with an obliterated sense swam in a similar pattern but to a reduced extent. The horizontal and vertical speeds of the intact fish were faster than those of the fish with a destroyed sense. Amplitude of vertical movements of the intact fish stretched from surface to bottom, while fish with destroyed vision tended towards midwater. Fish without an olfactory sense tended to be at the surface or near the bottom. Regular vertical movement would be effective in refreshing olfactory epithelia which might become acclimated when exposed to one odor. The vertical movements can be found in the horizontal zigzag movements in coastal and near shore migrations, as well as in the stream phase of homing. The zigzag movements, both horizontal and vertical, allow the fish to sense the water masses and locate the correct tributary.     

A blue protein as an inactivating factor for nitrite reductase from Alcaligenes faecalis strain S-6

A blue protein with a molecule weight of 12,000 containing 1 atom of type I Cu2+ was purified and crystallized from a denitrifying bacterium, Alcaligenes faecalis strain S-6, as an inactivating factor for copper-containing nitrite reductase of the same organism. Inactivation of the enzyme occurred when the enzyme was incubated aerobically with a catalytic amount of the blue protein in the presence of reducing agents such as cysteine and ascorbate. The blue protein acts as a direct electron donor for the enzyme to catalyze the reduction of nitrite, but in the absence of nitrite, the enzyme-reduced blue protein system reacts with oxygen to produce H2O2. A suicide inactivation mechanism of the enzyme due to this H2O2 production is proposed.     

Evidence of the Insensitivity of the alpha-inc Allele to the Function of the Homothallic Genes in Saccharomyces Yeasts

The possible function of the α-inc allele (an α mating-type allele that is insensitive to the function of the homothallic gene system) was investigated by means of protoplast fusion. The fusion of protoplasts prepared from haploid strains of α-inc HO HMα HMa and α ho hmα HMa gave rise mainly to nonmating clones (58 of 64 isolates) and a few clones (six of 64 isolates) showing α mating type. Thirty of the 58 nonmating clones showed the diploid cell size and 28 clones had a larger cell size. Tetrad analysis of the nonmating clones with diploid cell size indicated that they were a/α-inc diploid; the normal α allele in α/α-inc cells was preferentially switched to an a allele. This observation further indicated that the HO/ho HMα/hmα HMa/HMa genotype is effective for the conversion of the α to a and that the inconvertibility of the α-inc allele is due to the insensitivity of the mating-type allele to the functional combination of the homothallic genes. It was suspected that fusion products larger than diploid cells might have been caused by multiple fusion of protoplasts.