A Barley Efflux Transporter Operates in a Na+-Dependent Manner, as Revealed by a Multidisciplinary Platform.

Plant growth and survival depend upon the activity of membrane transporters that control the movement and distribution of solutes into, around, and out of plants. Although many plant transporters are known, their intrinsic properties make them difficult to study. In barley (Hordeum vulgare), the root anion-permeable transporter Bot1 plays a key role in tolerance to high soil boron, facilitating the efflux of borate from cells. However, its three-dimensional structure is unavailable and the molecular basis of its permeation function is unknown. Using an integrative platform of computational, biophysical, and biochemical tools as well as molecular biology, electrophysiology, and bioinformatics, we provide insight into the origin of transport function of Bot1. An atomistic model, supported by atomic force microscopy measurements, reveals that the protein folds into 13 transmembrane-spanning and five cytoplasmic α-helices. We predict a trimeric assembly of Bot1 and the presence of a Na(+) ion binding site, located in the proximity of a pore that conducts anions. Patch-clamp electrophysiology of Bot1 detects Na(+)-dependent polyvalent anion transport in a Nernstian manner with channel-like characteristics. Using alanine scanning, molecular dynamics simulations, and transport measurements, we show that conductance by Bot1 is abolished by removal of the Na(+) ion binding site. Our data enhance the understanding of the permeation functions of Bot1.

Cationic liposome colloidal stability in the presence of guar derivatives suggests depletion interactions may be operative in artificial tears.

The colloidal stability of cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) liposomes was measured in the presence of guar, carboxymethyl guar, hydroxypropyl guar (HPG), and in mixtures of HPG with boric acid. Carboxymethyl guar induced DOTAP aggregation, presumably by bridging flocculation. The interaction with HPG-borate, an anionic polyelectrolyte with labile charge groups, depended on ionic strength. Without added salt, HPG-borate (pH 9.2) adsorbed on the liposomes and destabilized them. In contrast, in 0.1 M NaCl, HPG-borate did not adsorb onto the liposomes. HPG, HPG-borate, and guar induced depletion flocculation of the liposomes at high polymer concentration. Depletion flocculation may be an important mechanism when HPG is employed in artificial tear formulations.

Absorption of boron after mouthwash treatment with Bocosept.

Healthy volunteers and patients with gingivitis were treated locally with the boron-containing bacteriostatic agent, Bocosept. Blood levels and urinary excretion of boron were examined by a spectrophotometric method. Blood concentrations after a single mouthwash with Bocosept slightly exceeded those after intake of 200 g raisins or a bottle of red wine. The blood levels during a one-week course of treatment showed a low rate of boron accumulation. The highest concentration was about 0.3 microgram B/ml, a level which does not seem to involve any risk of boron poisoning. The small amount taken up after mouthwash treatment with Bocosept does not appear to represent absorption by the oral mucosa. It seems more likely that the uptake of boron takes place in the intestine after ingestion of residual amounts from the mouth.

Formation of rhamnogalacturonan II-borate dimer in pectin determines cell wall thickness of pumpkin tissue.

Boron (B) deficiency results in inhibition of pumpkin (Cucurbia moschata Duchesne) growth that is accompanied by swelling of the cell walls. Monomeric rhamnogalacturonan II (mRG-II) accounted for 80% to 90% of the total RG-II in B-deficient walls, whereas the borate ester cross-linked RG-II dimer (dRG-II-B) accounted for more than 80% of the RG-II in control plants. The results of glycosyl residue and glycosyl linkage composition analyses of the RG-II from control and B-deficient plants were similar. Thus, B deficiency does not alter the primary structure of RG-II. The addition of (10)B-enriched boric acid to B-deficient plants resulted within 5 h in the conversion of mRG-II to dRG-II-(10)B. The wall thickness of the (10)B-treated plants and control plants was similar. The formation and possible functions of a borate ester cross-linked RG-II in the cell walls are discussed.

The Maillard reaction in demineralized dentin in vitro.

The Maillard reaction between carbohydrate and protein has been proposed as a cause of the browning of carious lesions. The aim of the present investigation was to determine the occurrence of this reaction in bovine dentin collagen in vitro and to establish the effect of the reaction on the proteolytic degradation of bovine dentin collagen in vitro. Slices of demineralized bovine dentin were incubated with 0.2 M glucose or buffer for 10 weeks at 37 degrees C. The formation of initial (furosine) and advanced (pentosidine) products of the Maillard reaction in dentin exposed to glucose was confirmed by HPLC. After reduction with NaBH4 to prevent intermediate Maillard products from further reaction, slices were either degraded with collagenase for fluorescence measurement or incubated with trypsin or pepsin to assess enzymatic degradation. Fluorescence characteristic for the Maillard reaction increased in glucose-exposed slices. Degradation of collagen by pepsin, but not by trypsin, was greatly depressed following glucose pretreatment. This may indicate an altered sensitivity to proteolytic degradation; the Maillard reaction thus has a potential role in caries arrestment.